Antibiotic compounds

ABSTRACT

The present invention relates to carbapenems and provides a compound of the formula (I):                    
     or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof wherein: 
     R 1  is 1-hydroxyethyl, 1-fluoroethyl or hydroxymethyl; 
     R 2  is hydrogen or C 1-4 alkyl; 
     R 3  is hydrogen or C 1-4 alkyl; 
     R 4  and R 5  are the same or different and are selected from hydrogen, halo, cyano, C 1-4 alkyl, nitro, hydroxy, carboxy, C 1-4 alkoxy, C 1-4 alkoxycarbonyl, aminosulphonyl, C 1-4 alkylaminosulphonyl, di-C 1-4 -alkylaminosulphonyl, carbamoyl, C 1-4 alkylcarbamoyl, di-C 1-4  alkylcarbamoyl, trifluoromethyl, sulphonic acid, amino, C 1-4 alkylamino, di-C 1 4 alkylamino, C 1-4 alkanoylamino, C 1-4 alkanoyl(H-C 1-4 alkyl)amino, C 1-4 alkanesulphonamido and C 1-4 alkylS(O) n — wherein n is zero, one or two: 
     with the proviso that there is no hydroxy or carboxy substituent in a position ortho to the link to —NR 3 —. Processes for their preparation, intermediates in their preparation, their use as therapeutic agents and pharmaceutical compositions containing them.

This is a continuation of application Ser. No. 08/833,056 filed Apr. 3,1997, now U.S. Pat. No. 5,856,321, which is a divisional of Ser. No.08/462,037, filed on Jun. 5, 1995, now U.S. Pat. No. 5,652,233, which isa continuation of Ser. No. 08/129,056, now U.S. Pat. No. 5,478,820 whichis a 371 of PCT/G893/00217, filed on Feb. 2, 1993.

The present invention relates to carbapenems and in particular to suchcompounds containing a carboxy substituted phenyl group. This inventionfurther relates to processes for their preparation, to intermediates intheir preparation, to their use as therapeutic agents and topharmaceutical compositions containing them.

The compounds of this invention are antibiotics and can be used in thetreatment of any disease that is conventionally treated with antibioticsfor example in the treatment of bacterial infection in mammals includinghumans.

Carbapenems were first isolated from fermentation media in 1974 and werefound to have broad spectrum antibacterial activity. Since thisdiscovery substantial investigations have been made into new carbapenemderivatives and many hundreds of patents and scientific papers have beenpublished.

The first, and so far the only, carbapenem to be commercially marketedis imipenem (N-formimidoyl thienamycin). This compound has a broadspectrum of antibacterial activity.

The present invention provides compounds with a broad spectrum ofantibacterial activity including against both Gram positive andnegative, aerobic and anaerobic bacteria. They exhibit good stability tobeta-lactamases. In addition representative compounds of this inventionexhibit a very favourable duration of action.

The carbapenem derivatives referred to herein are named in accordancewith the generally accepted semi-systematic nomenclature:

Accordingly the present invention provides a compound of the formula(I):

or a pharmaceutically acceptable salt or in vivo hydrolysable esterthereof wherein:

R¹ is 1-hydroxyethyl, 1-fluoroethyl or hydroxymethyl;

R² is hydrogen or C₁₋₄alkyl;

R³ is hydrogen or C₁₋₄alkyl;

R⁴ and R⁵ are the same or different and are selected from hydrogen,halo, cyano, C₁₋₄alkyl, nitro, hydroxy, carboxy, C₁₋₄alkoxy,C₁₋₄alkoxycarbonyl, aminosulphonyl, C₁₋₄alkylaminosulphonyl,di-C₁₋₄-alkylaminosulphonyl, carbamoyl, C₁₋₄alkylcarbamoyl,di-C-₁₋₄-alkylcarbamoyl, trifluoromethyl, sulphonic acid, amino,C₁₋₄alkylamino, di-C₁₋₄alkylamino, C₁₋₄alkanoylamino,C₁₋₄alkanoyl(N—C₁₋₄alkyl)amino, C₁₋₄alkanesulphonamido and C₁₋₄alkylS(O)wherein n is zero, one or two:

with the proviso that there is no hydroxy or carboxy substituent in aposition ortho to the link to —NR³—.

Alkyl when used herein includes straight chain and branched chainsubstituents for example methyl, ethyl, n-propyl, isopropyl, n-butyl andisobutyl.

Preferably R¹ is 1-hydroxyethyl.

R² is hydrogen or C₁₋₄alkyl for example methyl, ethyl, n-propyl,isopropyl or n-butyl. Preferably R² is hydrogen or methyl and inparticular R² is methyl.

R³ is hydrogen or C₁₋₄alkyl for example methyl, ethyl, n-propyl,isopropyl or n-butyl. Preferably R³ is hydrogen.

R⁴ and R⁵ are the same or different and are selected from hydrogen; halofor example fluoro, bromo or chloro; cyano; C₁₋₄alkyl for exampleaethyl, ethyl, n-propyl, isopropyl or n-butyl; nitro; hydroxy; carboxy;C₁₋₄alkoxy for example methoxy or ethoxy; C₁₋₄alkoxycarbonyl for examplemethoxycarbonyl, ethoxycarbonyl and n-propoxycarbonyl; aminosulphonyl;C₁₋₄alkylaminosulphonyl for example methylaminosulphonyl andethylaminosulphonyl; di-C₁₋₄alkylamino-sulphonyl for exampledimethylaminosulphonyl, methylethylaminosulphonyl anddi-ethylaminosulphonyl; carbamoyl; C₁₋₄alkylcarbamoyl for examplemethylcarbamoyl or ethylcarbamoyl; di-C₁₋₄alkylcarbamoyl for exampledimethylcarbamoyl or diethylcarbamoyl; trifluoromethyl; sulphonic acid;amino; C₁₋₄alkylamino for example methylamino or ethylamino;di-C₁₋₄alkylamino for example dimethylamino or diethylamino;C₁₋₄alkanoylamino for example acetamido or propionamido;C₁₋₄alkanoyl(N—C₁₋₄alkyl)amino for example N-methylacetamido;C₁₋₄alkanesulphonamido for example methanesulphonamido; andC₁₋₄alkylS(O) for example methylthio, methylsulphinyl ormethylsulphonyl.

In a particular aspect a suitable class of compounds is that in which R⁴and R⁵ are the same or different and are selected from hydrogen, fluoro,chloro, hydroxy, carboxy, cyano, nitro, methyl, ethyl, methoxy, ethoxy,methoxycarbonyl, carbamoyl, methylcarbamoyl, dimethylcarbamoyl,trifluoromethyl, sulphonic acid, methylsulphinyl, methylsulphonyl,methanesulphonamido or acetamido.

R⁴ and R⁵ may both be other than hydrogen but, in general, it isparticularly preferred that at least one of R⁴ and R⁵ is hydrogen.

Particularly preferred compounds are those in which R⁴ is hydrogen,carboxy, fluoro, chloro, methyl, methoxy, cyano, sulphonic acid ormethoxycarbonyl and R⁵ is hydrogen.

The present invention covers all epimeric, diastereoisomeric andtautomeric forms of the compounds of the formula (I) wherein theabsolute stereochemistry at the 5-position is as illustrated in formula(I). When a bond is represented by a wedge, this indicates that in threedimensions the bond would be coming out of the paper and when a bond ishatched, this indicates that in three dimensions the bond would be goingback into the paper. The compounds of the formula (I) have a number ofother stereocentres, namely: within the group R¹ (when R¹ is1-hydroxyethyl or 1-fluoroethyl); at the 6-position; at the 1-position(when R² is C₁₋₄alkyl); and at the 2′ and 4′ positions in thepyrrolidine ring:

Preferred compounds are those in which the beta-lactam ring protons arein trans configuration with respect to one another. When R¹ is1-hydroxyethyl or 1-fluoroethyl it is preferred that the 8-substituenthas the R-configuration. Thus a preferred class of compounds is that ofthe formula (III):

and pharmaceutically acceptable salts and in vivo hydrolysable estersthereof, wherein R², R³, R⁴ and R⁵ are as hereinbefore defined.

When R² is C₁₋₄alkyl for example methyl it is preferred that thecompound is in the form of the 1R configuration.

Preferred compounds are those in which the pyrrolidine ring has thefollowing absolute stereochemistry at the 2′- and 4′-positions:

A preferred class of compounds of the present invention is that of theformula (IV):

and pharmaceutically acceptable salts and in vivo hydrolysable estersthereof wherein R³, R⁴, and R⁵ are as defined hereinbefore in formula(I).

Particularly preferred compounds within the formula (IV) are thosewherein R³ is hydrogen and R⁴ and R⁵ are the same or different and areselected from hydrogen, fluoro, chloro, hydroxy, carboxy, cyano, nitro,methyl, ethyl, methoxy, ethoxy, methoxycarbonyl, carbamoyl,methylcarbamoyl, dimethylcarbamoyl, methanesulphonyl, trifluoromethyl,sulphonic acid, methylsulphinyl, methanesulphonamido or acetamido.

Especially preferred compounds within the formula (IV) are those whereinR³ and R⁵ are both hydrogen and R⁴is hydrogen, carboxy, fluoro, chloro,methyl, methoxy, cyano, sulphonic acid or methoxycarbonyl.

Suitable pharmaceutically acceptable salts include acid addition saltssuch as hydrochloride, hydrobromide, citrate, maleate and salts formedwith phosphoric and sulphuric acid. In another aspect suitable salts arebase salts such as an alkali metal salt for example sodium or potassium,an alkaline earth metal salt for example calcium or magnesium, anorganic amine salt for example triethylamine, morpholine,N-methylpiperidine, N-ethylpiperidine, procaine, dibenzylamine,N,N-dibenzylethylamine or amino acids for example lysine. For theavoidance of doubt there may be one, two or three salt-forming cationsdependent on the number of carboxylic acid functions and the valency ofsaid cations.

Preferred pharmaceutically acceptable salts are sodium and potassiumsalts. However, to facilitate isolation of the salt during preparation,salts which are less soluble in the chosen solvent may be preferredwhether pharmaceutically acceptable or not.

In vivo hydrolysable esters are those pharmaceutically acceptable estersthat hydrolyse in the human body to produce the parent compound. Suchesters can be identified by administering, eg. intravenously to a testanimal, the compound under test and subsequently examining the testanimal's body fluids. Suitable in vivo hydrolysable esters for carboxyinclude C₁₋₆alkoxymethyl esters for example methoxymethyl,C₁₋₆alkanoyloxymethyl esters for example pivaloyloxymethyl, phthalidylesters, C₃₋₈cycloalkoxycarbonyloxy-C₁₋₆alkyl esters for example1-cyclohexyloxycarbonyloxyethyl; 1,3-dioxolen-2-onylmethyl esters forexample 5-methyl-1,3-dioxolen-2-onylmethyl; andC₁₋₆alkoxycarbonyloxyethyl esters for example 1-methoxycarbonyloxyethyland may be formed at any carboxy group in the compounds of thisinvention. Suitable in vivo hydrolysable ester forming groups forhydroxy include acetyl, propionyl, pivaloyl, C₁₋₄alkoxycarbonyl forexample ethoxycarbonyl and phenylacetyl.

Particular compounds of the present invention are:

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-hydroxyphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-4-chlorophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-6-chlorophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxyphenylcarbanoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-6-methanesulphonylphenyl-carbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-4-fluorophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-6-fluorophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-2,4-difluorophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3,4-dicarboxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-4-hydroxyphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-metbylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3,5-dicarboxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(2-carbamoyl-3-carboxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-4-carbamoylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-carbamoylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-acetamidophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-4-acetamidophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-methylsulphonamidophenyl-carbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-sulphophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-6-carbamoylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-2-diuethylaminocarbonylphenyl-carbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(5R,6S,8R,2′S,4′S)-2-(2-(3-carboxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)carbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-4-methylphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-methylphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-6-methylphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-2-methoxyphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-4-methoxyphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-methoxyphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-6-methoxyphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-4,6-dimethoxyphenylcarbanoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-methoxycarbonylphenyl-carbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-cyanophenylcarbanoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-trifluoromethylphenyl-carbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-4,6-difluorophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-6-methylsulphinylphenyl-carbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-methylsulphonylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-ethylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-fluorophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-6-cyanophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxyN′-methylphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

and pharmaceutically acceptable salts and in vivo hydrolysable estersthereof.

Preferred compounds of the present invention are:

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-methylphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-methoxyphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-6-methoxyphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-methoxycarbonylphenyl-carbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-cyanophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-6-chlorophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-4-fluorophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-6-fluorophenylcarbamoyl)-pyrrolidin-4-ylthio)-6- (1-hydroxyethyl) - 1-methylcarbapenem-3-carboxylic acid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3,4-dicarboxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3,5-dicarboxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-sulphophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid,

and pharmaceutically acceptable salts thereof.

In order to use a compound of the formula (I) or a pharmaceuticallyacceptable salt or in vivo hydrolysable ester thereof for thetherapeutic treatment of mammals including humans, in particular intreating infection, it is normally formulated in accordance withstandard pharmaceutical practice as a pharmaceutical composition.

Therefore in another aspect the present invention provides apharmaceutical composition which comprises a compound of the formula (I)or a pharmaceutically acceptable salt or in vivo hydrolysable esterthereof and a pharmaceutically acceptable carrier.

The pharmaceutical compositions of this invention may be administered instandard manner for the disease condition that it is desired to treat,for example by oral, rectal or parenteral administration. For thesepurposes the compounds of this invention may be formulated by meansknown in the art into the form of, for example, tablets, capsules,aqueous or oily solutions or suspensions, emulsions, dispersiblepowders, suppositories and sterile injectable aqueous or oily solutionsor suspensions.

The compounds of the present invention may be formulated as dry powderfilled vials, which may contain the compound of the present inventionalone or as a dry blended mixture. For example an acidic compound of thepresent invention may be dry blended with an alkali metal carbonate orbicarbonate. Freeze dried formulations of compounds of the presentinvention, alone or as a mixture with standard excipients, are possible.Standard excipients include structure formers, cryoprotectants and pHmodifiers, such as, mannitol, sorbitol, lactose, glucose, sodiumchloride, dextran, sucrose, maltose, gelatin, bovine serum albumin(BSA), glycine, mannose, ribose, polyvinylpyrrolidine (PVP), cellulosederivatives, glutamine, inositol, potassium glutamate, erythritol,serine and other amino acids and buffer agents e.g. disodium hydrogenphosphate and potassium citrate.

In addition to the compounds of the present invention the pharmaceuticalcomposition of this invention may also contain, or be co-administeredwith, one or more known drugs selected from other clinically usefulantibacterial agents (for example other beta-lactams oraminoglycosides), inhibitors of beta-lactanase (for example clavulanicacid), renal tubular blocking agents (e.g. probenecid) and inhibitors ofmetabolising enzymes (for example inhibitors of dehydropeptidases, forexample Z-2-acylamino-3-substituted propenoates such as cilastatin) andN-acylated amino acids (for example see EP-A-178911) which reduceadverse effects on the kidney.

A suitable pharmaceutical composition of this invention is one suitablefor oral administration in unit dosage form, for example a tablet orcapsule which contains between 100 mg and 1 g of the compound of thisinvention.

A preferred pharmaceutical composition of the invention is one suitablefor intravenous, subcutaneous or intramuscular injection, for example asterile injectable containing between 1 and 50% V/W of the compound ofthis invention.

Specific examples of compositions, which are constituted as a 1%solution in water, freeze dried and may be made up by adding 0.9%aqueous sodium chloride solution to give the required concentration,preferably 1 mg-10 mg/ml, are as follows:

Composition 1 Compound of Example 1 50 mg Composition 2 Compound ofExample 1 50 mg Glycine 31 mg

Further specific examples of compositions are as above, but where thecompound of example 1 is, replaced by any one of examples 2 to 37.

The pharmaceutical compositions of the invention will normally beadministered to man in order to combat infections caused by bacteria, inthe same general manner as that employed for imipenem due allowancebeing made in terms of dose levels for the potency and duration ofaction of the compound of the present invention relative to the clinicaluse of imipenem. Thus each patient will receive a daily intravenous,subcutaneous or intramuscular dose of 0.05 to 5 g, and preferably 0.1 to2.5 g, of the compound of this invention, the composition beingadministered 1 to 4 times per day, preferably 1 or 2 times a day. Theintravenous, subcutaneous and intramuscular dose may be given by meansof a bolus injection. Alternatively the intravenous dose may be given bycontinuous infusion over a period of time. Alternatively each patientwill receive a daily oral dose which is approximately equivalent to thedaily parenteral dose. Thus a suitable daily oral dose is 0.05 to 5 g ofthe compound of this invention, the composition being administered 1 to4 times per day.

In a further aspect the present invention provides a process forpreparing the compounds of the formula (I) or a pharmaceuticallyacceptable salt or in vivo hydrolysable ester thereof which processcomprises deprotecting a compound of the formula (V):

wherein R², R⁴ and R⁵ are as hereinbefore defined (R⁴ and R⁵ beingoptionally protected if appropriate); —COOR⁶ and —COOR⁷ are carboxy orprotected carboxy; R⁸ is a group R³ or an amino protecting group; R⁹ ishydrogen or an amino protecting group; and R¹⁰ is a group R¹, protected1-hydroxyethyl or protected hydroxymethyl; and wherein at least oneprotecting group is present; and thereinafter if necessary;

(i) forming a pharmaceutically acceptable salt,

(ii) esterifying to form an in vivo hydrolysable ester.

Protecting groups may in general be chosen from any of the groupsdescribed in the literature or known to the skilled chemist asappropriate for the protection of the group in question, and may beintroduced by conventional methods.

Protecting groups may be removed by any convenient method as describedin the literature or known to the skilled chemist as appropriate for theremoval of the protecting group in question, such methods being chosenso as to effect removal of the protecting group with minimum disturbanceof groups elsewhere in the molecule.

The compounds of the formula (V) are novel and form another aspect ofthe invention.

Specific examples of protecting groups are given below for the sake ofconvenience, in which “lower” signifies that the group to which it isapplied preferably has 1-4 carbon atoms. It will be understood thatthese examples are not exhaustive. Where specific examples of methodsfor the removal of protecting groups are given below these are similarlynot exhaustive. The use of protecting groups and methods of deprotectionnot specifically mentioned is of course within the scope of theinvention.

A carboxyl protecting group may be the residue of an ester-formingaliphatic or araliphatic alcohol or of an ester-forming silanol (thesaid alcohol or silanol preferably containing 1-20 carbon atoms).

Examples of carboxy protecting groups include straight or branched chain(1-12C)alkyl groups (eg isopropyl, t-butyl); lower alkoxy lower alkylgroups (eg methoxymethyl, ethoxymethyl, isobutoxymethyl); loveraliphatic acyloxy lover alkyl groups, (eg acetoxymethyl,propionyloxymethyl, butyryloxymethyl, pivaloyloxymethyl); loveralkoxycarbonyloxy lover alkyl groups (eg 1-methoxycarbonyloxyethyl,1-ethoxycarbonyloxyethyl); aryl lover alkyl groups (eg p-methoxybenzyl,o-nitrobenzyl, p-nitrobenzyl, benzhydryl and phthalidyl); tri(loveralkyl)silyl groups (eg trimethylsilyl and t-butyldimethylsilyl);tri(lover alkyl)silyl lover alkyl groups (eg trimethylsilylethyl); and(2-6C)alkenyl groups (eg allyl and vinylethyl).

Methods particularly appropriate for the removal of carboxyl protectinggroups include for example acid-, base-, metal- or enzymically-catalysedhydrolysis.

Examples of hydroxyl protecting groups include lover alkenyl groups (egallyl); lover alkanoyl groups (eg acetyl); lover alkoxycarbonyl groups(eg t-butoxycarbonyl); lower alkenyloxycarbonyl groups (egallyloxycarbonyl); aryl lover alkoxycarbonyl groups (egbenzoyloxycarbonyl, p-methoxybenzyloxycarbonyl,o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl); tri loveralkylsilyl (eg trimethylsilyl, t-butyldimethylsilyl) and aryl loveralkyl (eg benzyl) groups.

Examples of amino protecting groups include formyl, aralkyl groups (egbenzyl and substituted benzyl, eg p-methoxybenzyl, nitrobenzyl and2,4-dimethoxybenzyl, and triphenylmethyl); di-p-anisylmethyl andfurylmethyl groups; lower alkoxycarbonyl (eg t-butoxycarbonyl); loweralkenyloxycarbonyl (eg allyloxycarbonyl); aryl lower alkoxycarbonylgroups (eg benzyloxycarbonyl, p-methoxybenzyloxycarbonyl,o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl; trialkylsilyl (egtrimethylsilyl and t-butyldimethylsilyl); alkylidene (eg methylidene);benzylidene and substituted benzylidene groups.

Methods appropriate for removal of hydroxy and amino protecting groupsinclude, for example, acid-, base-, metal- or enzymically-catalysedhydrolysis, for groups such as p-nitrobenzyloxycarbonyl, hydrogenationand for groups such as o-nitrobenzyloxycarbonyl, photolytically.

Preferred protecting groups for carboxy and hydroxy groups in compoundsof the formula (I) are the groups allyl and p-nitrobenzyl. A preferredmethod for removal of the allyl group is by palladium catalysis usingtetrakis(triphenylphosphine)palladium and Heldrum's acid, in a dipolaraprotic solvent tetrahydrofuran mixture, such asdimethylsulphoxide/tetrahydrofuran or1,3-dimethyl-2-oxo-tetrahydro-pyrimidine/tetrahydrofuran, or analcohol/tetrahydrofuran mixture such as isopropanol/tetrahydrofuran orethanol/tetrahydrofuran, preferably at ambient temperature.Alternatively, methylaniline nay be used in place of Heldrum's acid, indichloromethane. These conditions allow isolation of the product byprecipitation of the sodium salt on the addition of a sodium salt suchas sodium 2-ethylhexanoate.

A preferred method for removal of the p-nitrobenzyl group ishydrogenation using a palladium catalyst.

In another aspect of the present invention the compounds of the formulae(I) and (V) may be prepared by

a) reacting compounds of the formulae (VI) and (VII):

wherein R², R⁴-R¹⁰ are as hereinbefore defined and L is a leaving group,or

b) cyclising a compound of the formula (VIII):

wherein R², R⁴-R¹⁰ as hereinbefore defined and R¹ -R¹³ are independentlyselected from C₁₋₆-alkoxy, aryloxy, di-C₁₋₆alkylamino and diarylamino orany two of R¹¹-R¹³ represent o-phenylenedioxy; or one of R¹¹-R¹³ isC₁₋₄alkyl, allyl, benzyl or phenyl, and the other two values areindependently selected from C₁₋₄alkyl, trifluoromethyl or phenyl,wherein any phenyl group is optionally substituted with C₁₋₃alkyl orC₁₋₃alkoxy:

and wherein any functional group is optionally protected andthereinafter if necessary:

(i) removing any protecting groups;

(ii) forming a pharmaceutically acceptable salt;

(iii) esterifying to form an in vivo hydrolysable ester.

Suitably in the compound of the formula (VI) L is the reactive ester ofa hydroxy group such as a sulphonate (for exampleC₁₋₆alkanesulphonyloxy, trifluoromethanesulphonyloxy,benzenesulphonyloxy, toluenesulphonyloxy), a phosphoric ester (forexample a diarylphosphoric ester such as diphenylphosphoric ester) or Lis a halide (for example chloride). In an alternative L is a sulphoxidefor example —SOCH═CH—NHCOCH3 which may be readily displaced. PreferablyL is diphenylphosphoric ester (—OP(O)(OPh)₂).

Compounds of the formula (VI) and their preparation are well known inthe carbapenem literature, for example see EP-A-126587, EP-A-160391,EP-A-243686 and EP-A-343499.

The reaction between the compounds of the formulae (VI) and (VII) istypically performed in the presence of a base such as an organic aminefor example di-isopropylethylamine or an inorganic base for example analkali metal carbonate such as potassium carbonate. The reaction isconveniently performed at a temperature between −25° C. and ambient,suitably at about −20° C. The reaction is generally performed in anorganic solvent such as acetonitrile or dimethylformamide. The reactionis generally performed in a manner similar to that described in theliterature for similar reactions.

The compounds of the formula (VII) are novel and form another aspect ofthe present invention.

The compounds of the formula (VII) may be prepared by the deprotectionof a compound of the formula (IX):

wherein R⁴-R⁶, R⁸ and R⁹ as hereinbefore defined and R¹⁴ is a protectinggroup, for example C₁₋₆alkanoyl, C₁₋₆alkoxycarbonyl or benzoyl.Preferred values for R¹⁴ are acetyl and t-butoxycarbonyl. The compoundsof the formula (IX) can be converted to the compounds of the formula(VII) by standard methods of deprotection, for example acetyl groups canbe removed by basic hydrolysis in aqueous alkanol or alkenol for exampleallyl alcohol.

The compounds of the formula (IX) are novel and form another aspect ofthe present invention.

The compounds of the formula (IX) may be prepared by the reaction of anactivated derivative of a compound of the formula (X), which may beformed in situ, with a compound of the formula (XI):

wherein R⁴-R⁶, R⁸, R⁹ and R¹⁴ are as hereinbefore defined. Activatedderivatives of the compound of the formula (X) include acid halides,anhydrides and ‘activated’ esters such as 1H-benzo[1,2,3] triazol-1-yl,pentafluorophenyl and 2,4,5-trichlorophenyl esters or thebenzimidazol-2-yl ester of the thiocarboxylic acid corresponding to (X).The reaction of the compounds of the formulae (X) and (XI) is performedunder standard methods, for example in the presence of Vilsmeier reagent(thus forming the reactive derivative of (X) in situ) at temperatures inthe region −30° C. to 25° C. preferably in the region −20° C. to 5° C.

The compounds of the formulae (X) and (XI) are prepared by standardmethods known to the skilled chemist such as the methods of the Exampleshereinafter, the methods described in EP-A-126587 or by methodsanalogous or similar thereto.

Suitably, in the compounds of the formula (VIII), R¹¹-R¹³ areindependently selected from C1-6 alkoxy such as methoxy, ethoxy,isopropoxy, n-propoxy or n-butoxy; aryloxy such as optionally phenoxy;di-C₁₋₆alkylamino such as dimethylamino or diethylamino; diarylaminosuch as diphenylamino or any two of R¹¹-R¹³ represent o-phenylenedioxy.Preferably each of R¹¹-R¹³ have the same value and are C₁₋₆alkoxy forexample methoxy, ethoxy, isopropoxy or n-butoxy or are phenoxy.

The compounds of the formula (VIII) are cyclized under conventionalconditions known in the art to form compounds of the formula (V).Typical conditions are heating in a substantially inert organic solventsuch as toluene, xylene or ethyl acetate at temperatures in the region60-150° C. Typically the reaction is performed in an atmosphere ofnitrogen and is carried out in the presence of a radical scavenger forexample hydroquinone.

The compounds of the formula (VIII) may be formed and cyclized in situ.The compounds of the formula (VIII) may conveniently be prepared byreacting compounds of the formulae (XII) and (XIII):

wherein R², and R⁴-R¹³ are as hereinbefore defined. Suitably thecompound of the formula (XIII) is a phosphite or is the functionalequivalent of such a compound.

The reaction between the compounds of the formulae (XII) and (XIII) isconveniently performed in an organic solvent such as toluene, xylene,ethyl acetate, chloroform, dichloromethane, acetonitrile ordimethylformamide. Typically the reaction is carried out at an elevatedtemperature for example 60-150° C.

The compounds of the formula (XII) may be prepared by a number ofmethods known in the art. For example the compounds of the formula (XII)may be prepared by the acylation of a compound of the formula (XIV):

wherein R², R⁴-R⁶, and R⁸-R¹⁰ are as hereinbefore defined with acompound of the formula (XV):

Cl—Co—COOR⁷  (XV)

wherein R⁷ is as hereinbefore defined.

The compounds of the formula (XIV) may be prepared by reacting compoundsof the formulae (XVI) and (VII):

wherein R² and R¹⁰ are as hereinbefore defined. The compounds of theformula (XVI) are known in the art and may be reacted with the compoundsof the formula (VII) under conventional acylation methods known in theart.

Compounds of the formulae (VII), (XII) and (XIV) are novel and, as such,form another aspect of this invention.

The following biological test methods, data and Examples serve toillustrate the present invention.

Antibacterial Activity

The pharmaceutically acceptable carbapenem-compounds of the presentinvention are useful antibacterial agents having a broad spectrum ofactivity in vitro against standard laboratory microorganisms, bothGram-negative and Gram-positive, which are used to screen for activityagainst pathogenic bacteria. The antibacterial spectrum and potency of aparticular compound may be determined in a standard test system. Inparticular the carbapenems of the present invention show good stabilityto beta-lactamases and have a particularly good elimination half life inmammals. In general compounds show significant improvement overimipenem.

The antibacterial properties of the compounds of the invention may alsobe demonstrated in vivo in conventional tests.

Carbapenem compounds have generally been found to be relativelynon-toxic to warm-blooded animals, and this generalisation holds truefor the compounds of the present invention. Compounds representative ofthe present invention were administered to mice at doses in excess ofthose required to afford protection against bacterial infections, and noovert toxic symptoms or side effects attributable to the administeredcompounds were noted.

The following results were obtained for representative compounds on astandard in vitro test system using Diagnostic Sensitivity Test. Theantibacterial activity is described in terms of the minimum inhibitoryconcentration (HIC) determined by the agar-dilution technique with aninoculum size of 10⁴ CFU/spot.

MIC (μg/ml) EXAMPLE ORGANISM 1 2 4 10 ceftriaxone Enterobacter 0.06 0.030.03 0.01 0.06 cloacae 029 Enterobacter 1.00 1.00 0.50 0.25 32 cloacae108 E. coli 0.03 0.02 0.02 0.01 0.03 TEM S. aureus 0.25 0.50 0.25 0.132.0 147N

In the examples:

(a) NNR spectra were taken at 200 MHz or 400 MHz ;

(b) Allyloxy means the propen-1-yloxy group —OCH₂CH═CH₂;

(c) THF means tetrahydrofuran;

(d) DMF means dimethylformamide;

(e) Heldrum's acid is 2,2-dimethyl-1,3-dioxane-4,6-dione.

(f) Evaporation of solvents was carried out under reduced pressure;

(g) EtOAc means ethyl acetate;

(h) EEDQ means N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline;

(i) DMSO means dimethyl sulfoxide;

(j) DCCI means dicyclohexylcarbodiimide; and

(k) The peak positions in NXR spectra taken in DMSO-d₆ and aceticacid-d₄ vary depending on the ratio of DMSO to acetic acid.

EXAMPLE 1

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-5-hydroxyphenylcarbamoyl)-pyrrolidin-4-yl-thio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt

To a solution of allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxy-carbonyl-2-(3-allyloxy-5-allyloxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate(500 mg, 0.72 mM) and 2,2-dimethyl-1,3-dioxane-4,6-dione (Heldrum'sacid, 829 mg, 5.75 mM) in a mixture of DMF (8 ml) and THF (4 ml), underan argon atmosphere, was added tetrakis(triphenylphosphine)palladium (83mg, 0.072 mM). The solution was stirred, under argon with protectionfrom light, for 2 hours. The solution was diluted with diethyl ether (40ml), and the resultant precipitate centrifuged, and the supernatant wasremoved. The product was washed by resuspension in ether followed bycentrifugation, and finally dried under high vacuum. The crude productwas dissolved in water (10 ml) and the pH adjusted to 6.8 with NaHCO₃After filtration, the solution was subjected to chromatography on DiaionCHP20P resin, and the fractions combined as appropriate to give thetitle product (66%). Nmr (DMSO-d₆+acetic acid-d₄): δ 1.18 (d, 6H); 1.82(m, part obscured, 1H); 2.79 (m, 1H); 3.03 (dd, 1H); 3.22 (dd, 1H); 3.38(quintet, 1H); 3.57 (dd, 1H); 3.82 (quintet, 1H); 3.99 (quintet, 1H);4.19 (dd+m, 2H); 7.13 (t, 1H); 7.44 (t, 1H); 7.65 (t, 1H).

The starting materials were prepared as follows:

Allyl 3-allyloxy-5-aminobenzoate

3-Hydroxy-5-nitrobenzoic acid (3.9 g, 21.3 mM) was dissolved in DMF (55ml), and anhydrous K₂CO₃ (11.78 g, 76.5 mM) added with stirring. Allylbromide (5.4 ml, 62.4 mM) was run in, and the mixture stirred for 18hours at ambient temperature. The solvent was removed by evaporation,the residue treated with water, the pH was adjusted to 5.5, and productwas extracted into ethyl acetate. The combined extracts were washed withaqueous NaH₂PO₄, water, brine, and dried over MgSO₄. The residue afterevaporation was subjected to chromatography on silica, eluting with amixture of petrol/EtOAc (10:1), to give allyl 3-allyloxy-5-nitrobenzoate(5.94 g, 90%). Nmr (CDCl₃): δ 4.66 (dt, 2H); 4.87 (dt, 2H); 5.31-5.52(m, 4H); 5.94-6.14 (m, 2H); 7.92 (m, 2H); 8.46 (t, 1H).

Ms (CI): 264 (MH)⁺

The above ester (2 g, 7.6 mM) was dissolved in ethyl acetate (15 ml),and added to a suspension of SnCl₂.2H₂O(13.7 g, 61 mM), heated underreflux, in ethyl acetate (35 ml) under argon. The mixture was heated toreflux for 4 hours, cooled, and poured into a mix of 880 ammonia (20 ml)and water (20 ml). The organic layer was separated and three furtherextractions made with ethyl acetate. The combined extracts were washedwith dilute ammonia solution, water and brine, dried over MgSO₄, andevaporated to give a yellow oil of allyl 3-allyloxy-5-aminobenzoate(1.53 g, 86%). Nmr (CDCl₃): δ 3.60 (br, 2H); 4.53 (dt, 2H); 4.78 (dt,2H); 5.25-5.44 (m, 4H); 5.96-6.12 (m, 2H); 6.43 (dt, 1H); 7.00 (m, 2H).

Ms (CI): 233 (NH)⁺

Preparation of Side Chain Pyrrolidin-4-ylthioacetate

The cyclohexylamine salt of4-acetylthio-1-allyloxycarbonyl-2-carboxy-pyrrolidine (5.6 g, 15 mM) wassuspended in ethyl acetate, and shaken successively with 2M HCl (20 mland 10 ml), water and brine, and the ethyl acetate layer dried overMgSO₄. Evaporation gave the free acid. Vilsmeier reagent was prepared bytreatment of dimethylformamide (0.51 ml, 6.6 mM) in dichloromethane (20ml) under argon with oxalyl chloride (0.52 ml, 6 mM) in dichloromethane(5 ml) for 30 minutes.4-Acetylthio-1-allyloxycarbonyl-2-carboxypyrrolidine (1.64 g, 6 mM) indichloromethane (7 ml) was added to this in one portion, followed byN-methylmorpholine (0.79 ml, 7.2 mM), in dichloro-methane (3 ml) andstirring continued for 30 minutes at −10°. After cooling to −20°, allyl3-allyloxy-5-aminobenzoate (1.39 g, 5.9 mM) plus N-methylmorpholine(0.79 ml, 7.2 mM) dissolved in dichloromethane (15 ml) were addeddropvise. The temperature was allowed to rise to 0°, and the reactionstored for 18 hours. After dilution with dichloromethane (100 ml), themixture was washed with 2H HCl, H₂O, and saturated NaHCO₃, dried overMgSO₄, and evaporated. Crude material was purified by medium pressurechromatography on silica using a gradient of petrol in dichloromethane(3:1 to 2:1) to give(2S,4S)-1-allyloxy-carbonyl-2-(3-allyloxy-5-allyloxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthioacetateas a gum (2.37 g, 81%). Nmr (CDCl₃): δ 2.32 (s, 3H); 2.58 (br, 2H); 3.39(dd, 1H); 4.03 (quintet, 1H); 4.13 (dd, 1H); 4.55 (t, part obscured,1H); 4.58 (dt, 2H) 4.68 (dt, 2H); 4.81 (dt, 2H); 5.23-5.49 (m, 6H);5.84-6.15 (m, 3H); 7.36 (t, 1H); 7.57 (t, 1H); 7.66 (t, 1H); 9.10 (br,1H).

Ms (+ve FAB): 489 (MH)⁺, 511 (M+Na)⁺

Conversion to Pyrrolidin-4-ylthiols

(2S,4S)-4-Acetylthio-1-allyloxycarbonyl-2-(3-allyloxy-5-allyloxycarbonylphenylcarbamoyl)pyrrolidine(1.89 g, 3.9 mM) was dissolved in allyl alcohol (25 ml) and the solutionflushed with argon. 1H Sodium hydroxide (4 ml, 4 mM) was added, themixture stirred at ambient temperature for 2 hours, and then evaporatedto dryness. The residue taken up in ethyl acetate (100 ml), washed with2M HCl, water, NaHCO₃, brine, dried (MgSO₄) and evaporated, to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxy-5-allyloxycarbonylphenylcarbamoyl)-pyrrolidin-4-ylthiolas a gum (1.57 g, 76%). Crude material was used in the next stage.

Preparation of Protected Carbapenems

A solution of allyl(1R,5R,6S,8R)-6-(1-hydroxyethyl)-1-methyl-2-diphenylphosphoryloxycarbapenem-3-carboxylate(1.5 g, 3 mM) was dissolved in dry acetonitrile (18 ml) under argon,cooled to −20°, and diisopropylethylamine (0.63 ml, 3.6 mM) added,followed by dropwise addition of(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxy-5-allyloxy-carbonylphenylcarbamoyl)pyrrolidin-4-yl-thiol(1.57 g, 3.5 mm) in acetonitrile (12 ml). The reaction mixture was thenstored at −20° for 3 days. Solvent was evaporated, and the residuepurified by medium pressure chromatography on silica with gradientelution (dichloromethane/ethyl acetate 40:60 to 70:30), to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyloxy-5-allyloxy-carbonylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylateas a gum (1.25 g, 60%). Nmr (CDCl₃): δ 1.23 (d, 3H); 1.35 (d, 3H); 2.65(br, 2H); 3.26,3.31 (dd overlapping m, 2H); 3.46 (m, 1H); 3.79 (quintet,1H); 4.01 (dd, 1H); 4.19-4.29 (m, 2H); 4.50-4.78 (m, 9H); 5.19-5.46 (m,8H); 5.83-6.12 (m, 4H); 7.36 (br S, 1H); 7.64 (m, 2H); 9.00 (br, 1H).

Ms (+ve FAB): 696 (MH) , 718 (M+Na)⁺

Allyl(1R,5R,6S,8R)-6-(1-hydroxyethyl)-1-methyl-2-diphenyl-phosphoryloxy-carbapenem-3-carboxylatewas prepared as follows.

To a solution of allyl(1R,5S,6S,8R)-6-(1-hydroxyethyl)-1-methyl-2-oxocarbapenam-3-carboxylate(2.66 mmol) [prepared in situ from allyl2-diazo-3-oxo-4-methyl-4-(3-(1-hydroxyethyl)-2-oxoazetidin-4-yl)butanoateand rhodium octanoate: see for example EP-A-208889] anddiisopropylethylamine (1.1 equivalents in acetonitrile, at 0° C., underan argon atmosphere, was added dropwise diphenyl chlorophosphate (1.1equivalents). The solution was stirred at ambient temperature for 30minutes to form the corresponding 2-diphenylphosphoryloxycarbapenem.

The following further examples were prepared:

POSITION OF PHENYL SUBSTITUENT Example 2 3 4 5 6 R¹ R²  2 H COOH Cl H HMe H  3 H COOH H H Cl Me H  4 H COOH H H H Me H  5 H COOH H H SO₂CH₃ MeH  6 H COOH F H H Me H  7 H COOH H H F Me H  8 F COOH F H H Me H  9 HCOOH COOH H H Me H 10 H COOH OH H H Me H 11 H COOH H COOH H Me H 12 HCOOH H H H Me H 13 H COOH H CONH₂ H Me H 14 H COOH H H CONH₂ Me H 15CONMe₂ COOH H H H Me H 16 H COOH H NHCOCH₃ H Me H 17 H COOH NHCOCH₃ H HMe H 18 H COOH H NHSO₂Me H Me H 19 H COOH H SO₃H H Me H 20 H COOH H H HH H 21 H COOH H CN H Me H 22 H COOH H OMe H Me H 23 H COOH H H SOMe Me H24 H COOH H SO₂Me H Me H 25 H COOH H CF₃ H Me H 26 H COOH H H OMe Me H27 H COOH OMe H H Me H 28 OMe COOH H H H Me H 29 H COOH H H Me Me H 30 HCOOH Me H H Me H 31 H COOH H Me H Me H 32 H COOH H —COOMe H Me H 33 HCOOH F H F Me H 34 H COOH OMe H OMe Me H 35 H COOH H H CN Me H 36 H COOHH F H Me H 37 H COOH H H H Me Me

EXAMPLE 2

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-4-chlorophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared in the same general manner as Example 1except that after the addition of the palladium catalyst the solutionwas gently warmed to dissolve the catalyst and was stirred under argonwith protection from light for 1 hour. A solution of sodium2-ethylhexanoate in THF was added and the combined solutions poured intoTHF with vigorous stirring. The resultant precipitate was centrifugedand supernatant removed. The product was washed twice by resuspension inTHF followed by centrifugation and finally dried under high vacuum togive the title product. Nmr (DMSO-d₆+acetic acid-d₄): δ 1.17 (d, 6H);1.85 (m, obscured, 1H); 2.73 (m, obscured, 1H); 2.95 (dd, 1H); 3.21 (dd,1H); 3.40 (m, 1H); 3.54 (dd, 1H); 3.78 (quintet, 1H); 3.99 (t, 1H); 4.11(t, 1H); 4.18 (dd, 1H); 7.41 (d, 1H); 7.75 (dd, 1H); 8.06 (d, 1H).

Ms (+ve FAB): 532/534 (MH)⁺, (Na salt)⁺; 554/556 (Na₂ salt)⁺

The starting materials were prepared as follows:

2-Chloro-5-nitrobenzoic acid was allylated essentially as in Example 1,except that the final extraction solvent was toluene, to give allyl2-chloro-5-nitrobenzoate. Nmr (CDCl₃): δ 4.89 (dt, 2H); 5.33-5.51 (m,2H); 5.96-6.15 (m, 1H); 7.66 (d, 1H); 8.27 (dd, 1H); 8.72 (d, 1H).

Ms (CI): 241/243 M⁺, 259/261 (M+NH₄)⁺

Stannous chloride dihydrate was refluxed in ethanol, under an argonblanket, to give a solution. The heat was removed, and the above nitrocompound in ethanol was run in. Refluxing was then continued for 3hours, the mixture cooled, and solvents removed. The residue wasdissolved in ethyl acetate, and treated with 880 ammonia until basic.The organic phase was decanted from precipitated tin salts, and theslurry re-extracted similarly with more solvent. Combined organic phaseswere then washed with diluted ammonia, water, and brine, before dryingover MgSO₄. Evaporation gave allyl 5-amino-2-chlorobenzoate. Nmr(CDCl₃): δ 3.74 (br, 2H); 4.81 (dt, 2H); 5.27-5.47 (m, 2H); 5.93-6.13(m, 1H); 6.73 (dd, 1H); 7.15 (d, 1H); 7.24 (d, 1H).

Ms (CI): 212/214 M⁺, 229/231 (M+NH₄)⁺

The above amine was condensed with proline acid as Example 1, purifyingby chromatography using a gradient of dichloromethane/diethyl ether(100:0 to 95:5) to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-4-chlorophenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.33 (s, 3H); 2.57 (br, 2H); 3.39 (dd, 1H); 4.03(quintet, 1H); 4.13 (dd, 1H); 4.55 (t, 1H); 4.66 (dt, 2H)) 4.83 (dt,2H); 5.24-5.47 (m, 4H); 5.85-6.02 (m, 2H); 7.36 (d, 1H); 7.70 (dd, 1H);7.92 (d, 1H); 9.32 (br, 1H).

Ms (+ve FAB): 467/469 (MH)⁺, 489/491 (M+Na)⁺

The above thioacetate was deacetylated to thiol, and condensed withcarbapenem phosphate as in Example 1, purifying by chromatography usinggradient elution from dichioromethane to ethyl acetate to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyloxycarbonyl-4-chlorophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate. Nmr (CDCl₃) δ1.24 (d, 3H); 1.36 (d, 3H); 2.65(br, 2H); 3.25 (dd overlapping m, 2H); 3.48 (m, 1H); 3.80 (quintet, 1H);3.98 (dd, 1H); 4.20-4.31 (dd overlapping quintet, 2H); 4.52 (t, 1H);4.51-4.76 (m, 4H); 4.83 (dt, 2H); 5.20-5.47 (m, 6H); 5.85-6.11 (m, 3H);7.39 (d, 1H); 7.77 (dd, 1H); 7.99 (d, 1H); 9.05 (br, 1H).

Ms (+ve FAB): 674/676 (MH)⁺, 696/698 (M+Na)⁺

EXAMPLE 3

(1R,5S,6S,8R,2′S,4′S)-2-(2-(5-Carboxy-2-chlorophenylcarbamoyl)-pyrrolidin-4-yl-thio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 2. Nmr(DMSO-d₆+acetic acid-d₄): δ 1.16 (d, 3H); 1.19 (d, 3H); 1.82 (in,obscured, 1H); 2.70 (dd overlapping m, 2H); 3.22 (dd, 1H); 3.35-3.60(overlapping m, 3H); 3.95-4.08 overlapping m, 2H); 4.16 (dd, 1H); 7.57(d, 1H); 7.69 (dd, 1H); 8.36 (d, 1H).

Ms (+ve FAB): 532/534 (MH)⁺, (Na salt)⁺; 554/556 (Na₂ salt)

The starting materials were prepared as follows:

4-Chloro-3-nitrobenzoic acid was allylated essentially as in Example 1above to give allyl 4-chloro-3-nitrobenzoate. Nmr (CDCl₃): δ 4.86 (d,2H); 5.31-5.48 (m, 2H); 5.94-6.13 (m, 1H); 7.50 (d, 1H); 8.18 (dd, 1H);8.52 (d, 1H).

Ms (CI): 241/243 M⁺, 259/261 ((M+NH₄)⁺

Reduction of the above nitro compound by the method of Example 2 gaveallyl 3-amino-4-chlorobenzoate. Nmr (CDCl₃): δ 4.08 (br, 2H); 4.79 (dt,2H); 5.25-5.44 (m, 2H); 5.92-6.11 (m, 1H); 7.30 (d, 1H); 7.38 (dd, 1H);7.47 (d, 1H).

Ms (CI): 212/214 M⁺, 229/231 ((M+NH₄)⁺

The above amine was condensed with proline acid as Example 1, purifyingby chromatography using a gradient of dichloromethane/diethyl ether(100:0 to 95:5) to give(2S,4S)-1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-chlorophenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.32 (s, 3H); 2.56 (br, 1H); 2.66 (br, 1H); 3.43 (dd,1H); 4.04 (quintet, 1H); 4.16 (dd, 1H); 4.61 (t, 1H); 4.66 (dt, 2H) 4.82(dt, 2H); 5.21-5.45 (m, 4H); 5.84-6.11 (m, 2H); 7.45 (d, 1H); 7.77 (dd,1H); 9.00 (d, 1H); 9.08 (br, 1H).

Ms (+ve FAB): 467/469 (MH)⁺, 489/491 (M+Na)⁺

The above thioacetate was deacetylated to thiol, and condensed withcarbapenem phosphate as Example 1, purifying by chromatography usinggradient elution from dichloromethane to ethyl acetate to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-chlorophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methyl-carbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.23 (d, 3H); 1.36 (d, 3H); 2.65 (br, 2H); 3.24 (ddoverlapping m, 2H); 3.88 (quintet, 1H); 4.08 (1R, 1H); 4.19-4.30 (ddoverlapping quintet, 2H); 4.60 (t, 1H); 4.67 (a, 4H).; 4.82 (dt, 2H);5.18-5.45 (m, 6H); 5.82-6.01 (m, 3H); 7.44 (d, 1H); 7.76 (dd, 1H); 9.04(d, 1H); 8.98 (br, 1H).

Ms (+ve FAB): 674/676 (MH)⁺, 696/698 (M+Na)⁺

EXAMPLE 4

(1R, 5S,6S, 8R,2 ′S,4′S)-2-(2-(3-Carboxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 2 exceptthat a mixture of DMSO and THF was used. Nmr (DMSO-d₆+acetic acid-d₄): δ1.18 (d, 6H); 1.94 (m, obscured, 1H); 2.85 (m, 1H); 3.10 (dd, 1H); 3.23(dd, 1H); 3.40 (quintet, 1H); 3.66 (dd, 1H); 3.89 (quintet, 1H); 3.99(t, 1H); 4.21 (dd, 1H); 4.27 (t, 1H); 7.46 (t, 1H); 7.71 (d, 1H); 7.86(d, 1H); 8.27 (s, 1H).

Ms (+ve FAB): 498 (MH)⁺, (Na salt)⁺; 520 (Na₂ salt)⁺

The starting materials were prepared as follows:

3-Nitrobenzoic acid was allylated essentially as in Example 1, exceptthat the final extraction solvent was diethyl ether, to give allyl3-nitrobenzoate. Nmr (CDCl₃): δ 4.88 (d, 2H); 5.33-5.49 (m, 2H);5.96-6.17 (m, 1H); 7.66 (t, 1H); 8.41 (td, 2H); 8.88 (t, 1H).

Reduction of the above nitro compound by the method of Example 2, exceptthat the solvent was methanol, gave allyl 3-aminobenzoate. Nmr (CDCl₃) δ3.38 (br, 2H); 4.79 (dt, 2H); 5.24-5.44 (m, 2H); 5.93-6.09 (m, 1H); 6.86(dm, 1H); 7.21 (t, 1H); 7.37 (t, 1H); 7.45 (dt, 1H).

Preparation of Side Chain Pyrrolidin-4-ylthioacetate

(2S,4S) -4-Acetylthio-1-allyloxycarbonyl-2-carboxypyrrolidine (2.54 g,9.3 mM), allyl 3-aminobenzoate (1.5 g, 8.5 mM), and2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (2.72 g, 11 mM) weredissolved in toluene (50 ml) and stirred for 18 hours at ambienttemperature. The reaction mixture was diluted with ethyl acetate (150ml) and washed with 2M HCl (3 by 30 ml), water, saturated NaHCO₃, andbrine. Drying over MgSO₄ and evaporation gave(2S,4S)-4-acetylthio-1-allyloxycarbonyl-2-(3-allyloxycarbonylphenylcarbamoyl)pyrrolidineas a gum (3.7 g, 100%) in a state sufficiently pure for further work.Nmr (CDCl₃): δ 2.32 (s, 3H); 2.60 (br, 2H); 3.40 (dd, 1H); 4.03(quintet, 1H); 4.13 (dd, 1H); 4.57 (t, 1H); 4.66 (dm, 2H); 4.82 (dt,2H); 5.23-5.46 (m, 4H); 5.86-6.12 (m, 2H); 7.41 (t, 1H); 7.82 (d, 1H);7.91 (d, 1H); 8.07 (t, 1H); 9.18 (br, 1H).

The above thioacetate was deacetylated to thiol, and condensed withcarbapenem phosphate as Example 1, purifying by chromatography usinggradient elution from dichloromethane to ethyl acetate/dichloromethane1:1, to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyloxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.24 (d, 3H); 1.36 (d, 3H); 2.64 (br, 2H); 3.26,3.28 (ddoverlapping m, 2H); 3.48 (m, 1H); 3.81 (quintet, 1H); 4.01 (dd, 1H);4.22-4.32 (m, 2H); 4.54 (t, 1H); 4.62-4.75 (m, 4H); 4.82 (m, 2H);5.19-5.45 (m, 8H); 5.82-6.10 (m, 4H); 7.41 (d, 1H); 7.81 (d, 1H); 7.92(dm, 1H); 8.11 (t, 1H); 8.98 (br, 1H).

Ms (+ve FAB): 640 (MH)⁺, 662 (M+Na)⁺

EXAMPLE 5

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-6-methanesulphonylphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid disodium salt was prepared from the appropriate protectedcarbapenem as described in Example 1. Nmr (DMSO-d₆+acetic acid-d₄): δ1.20 (d, 6H); 1.99 (quintet, 1H); 2.75 (m, part obscured, 1H); 2.87 (dd,1H); 3.22 (s, 3H); 3.25 (dd, part obscured, 1H); 3.44 (quintet, 1H);3.62 (dd, 1H); 3.75 (quintet, 1H); 4.03 (quintet, 1H); 4.16-4.23 (m,2H); 7.90 (dd, 1H); 8.17 (d, 1H); 9.00 (d, 1H).

Ms (+ve FAB): 554 (MH)⁺, 576 (Na salt)⁺, 598 (Na₂ salt)⁺

The starting materials were prepared as follows:

4-hethanesulphonyl-3-nitrobenzoic acid was allylated essentially as inExample 1, except that crude product was purified by chromatography oversilica, eluting with a gradient of dichloromethane todichloromethane/diethyl ether 9:1, to give allyl4-methanesulphonyl-3-nitrobenzoate. Nmr (CDCl₃): δ 3.45 (s, 3H); 4.90(dt, 2H); 5.30-5.49 (m, 2H); 5.96-6.12 (m, 1H); 8.29 (d, 1H); 8.40-8.46(m, 2H).

Reduction as in Example 1, except that the solvent was methanol, gaveallyl 3-amino-4-methanesulphonylbenzoate. Nmr (CDCl₃): δ 3.07 (s, 3H);4.82 (dt, 2H); 5.05 (br, 2H); 5.29-5.44 (m, 2H); 5.95-6.11 (m, 1H); 7.46(m, 2H); 7.81 (d, 1H).

Ms (+ve FAB): 256 (MH)⁺, 273 (M+NH₄)⁺

The above amine was condensed with proline acid as Example 1, purifyingby chromatography using a gradient of dichloromethane/diethyl ether(100:0 to 90:10), to give(2S,4S)-1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-methanesulphonylphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.31 (s, 3H); 2.41 (m, 1H); 2.80 (m, 1H); 3.11 (s, 3H);3.51 (dd, 1H); 4.00-4.18 (m, 2H); 4.53 (dd, 1H); 4.65 (2m, 2H) 4.87 (dt,2H); 5.23-5.47 (m, 4H); 5.83-6.13 (m, 2H); 7.93 (dd, 1H); 8.03 (d, 1H);9.09 (br s, 1H).

Ms (+ve FAB): 511 (MH)⁺, 533 (M+Na)⁺

The above thioacetate was deacetylated to thiol, and condensed withcarbapenem phosphate as Example 1, purifying by chromatography usinggradient elution from dichloromethane through to ethyl acetate to giveallyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-methanesulphonylphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.21 (d, 3H); 1.35 (d, 3H); 2.43 (m, 1H); 2.75 (br, 1H);3.08 (s, 3H); 3.23 (dd overlapping m, 2H); 3.55 (dd, 1H); 3.85-4.08 (m,2H); 4.19-4.28 (m, 2H); 4.53-4.68 (m, 5H); 4.86 (dt, 2H); 5.17-5.47 (m,6H); 5.79-6.12 (m, 3H); 7.92 (dd, 1H); 8.00 (d, 1H); 9.16 (br s, 1H);10.14 (br, 1H).

Ms (+ve FAB): 718 (MH)⁺, 740 (M+Na)

EXAMPLE 6

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-4-fluorophenylcarbamoyl)pyrrolidin-4-yl-thio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid

To a solution of allyl(1R,5S,6S,8R,2′S,4′S)-2-(1′-allyloxycarbonyl-2′-(3-allyloxycarbonyl-4-fluorophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate(199 mg, 0.3 mM) and 2,2-dimethyl-1,3-dioxane-4,6-dione (Heldrum's acid,259 mg, 1.8 mM) in DMF (1.5 ml), under an argon atmosphere, was addedtetrakis(triphenylphosphine)palladium (10 mg, 0.009 mM) in THF (0.1 ml).The solution was stirred under argon for 2 hours andtetrakis(triphenyl-phosphine)palladium (5 mg, 0.0045 mM) in THF (0.1 ml)was added. After stirring for 30 minutes, THF (3 ml) and ether (9 ml)were added, the resultant solid filtered off, washed with ether (9 ml),and dried under high vacuum to give the title product (72 mg, 49%). Nmr(DMSO-d₆+acetic acid-d₄): δ 1.30 (d, 6H); 2.00 (m, part obscured, 1H);2.91 (m, part obscured, 1H); 3.13 (dd, 1H); 3.36 (dd, 1H); 3.55 (dq,1H); 3.73 (dd, 1H); 3.95 (m, 1H); 4.12 (m, 1H); 4.29 (dd, 1H); 4.34 (dd,1H); 7.40 (dd, 1H); 7.98 (m, 1H); 8.32 (dd, 1H).

Ms (+ve FAB): 494 (MH)⁺, 516 (M+Na)⁺

The starting materials were prepared as follows:

Allyl 5-amino-2-fluorobenzoate

2-Fluoro-5-nitrobenzoic acid (4.16 g, 22.5 mM) was dissolved in DMF (45ml), and anhydrous K₂CO₃ (4.65 g, 33.7 mM) added with stirring. Allylbromide (2.38 ml, 28.1 mM) was run in, the mixture was stirred for 18hours at ambient temperature before being poured into water (450 ml) andextracted with diethyl ether (3×100 ml). The combined extracts weredried over MgSO₄, and evaporated to give a yellow oil (5.4 g). The oilwas purified by chromatography on silica, eluting with a mixture ofethyl acetate/hexane (12.5:87.5), to give allyl 2-fluoro-5-nitrobenzoate(4.64 g, 92%). Nmr (CDCl₃): δ 4.89 (d, 2H); 5.30-5.50 (m, 2H); 5.90-6.10(m, 1H); 7.32 (t, 1H); 8.38-8.46 (m, 1H); 8.86 (dd, 1H).

Ms (EI): 226 (MH)⁺; (CI): 225 M⁺, 243 (M+NH₄)⁺

The above ester (2.47 g, 10.97 mN) was dissolved in methanol (40 ml),and stannous chloride dihydrate (9.89 g, 43.76 mM) in conc HCl (9 ml)was added to the stirred solution while maintaining the temperaturebetween 5° and 15°. The mixture was then stirred overnight at ambienttemperature before being poured into water (200 ml) and neutralized withsolid NaHCO₃ (17.6 g) to pH6. The mixture was extracted with chloroform(3×200 ml), the combined extracts were dried over MgSO₄, and evaporatedto give allyl 5-amino-2-fluorobenzoate (2.09 g, 98%) as a yellow oil.Nmr (CDCl₃): δ 3.60 (br s, 2H); 4.82 (dt, 2H); 5.25-5.48 (m, 2H);5.93-6.12 (m, 1H); 6.78 (ddd, 1H); 6.93 (dd, 1H); 7.20 (dd, 1H).

Ms (EI): 195 M⁺; (CI): 196 (MH)⁺; 213 ((M+NH₄)⁺

The above amine was condensed with the proline acid as in Example 1. Theproduct was purified by chromatography on silica using ethylacetate/hexane (42.5:57.5) to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-4-fluorophenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.32 (s, 3H); 2.50-2.70 (br s, 2H); 3.40 (d, 1H);3.98-4.20 (m, 2H); 4.56 (t, 1H); 4.67 (dt, 2H); 4.84 (dt, 2H); 5.20-5.50(m, 4H); 5.83-6.12 (m, 2H); 7.10 (dd, 1H); 7.80-7.89 (m, 1H); 7.93 (dd,1H); 8.90-9.40 (br s, 1H).

Ms (+ve FAB): 451 (MH)⁺, 473 (M+Na)⁺

Conversion to Pyrrolidin-4-ylthiol

(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-4-fluorophenylcarbamoyl)-pyrrolidin-4-ylthioacetate (1.33 g, 2.96 mN) was dissolved inallyl alcohol (30 ml) and the solution flushed with argon. 1M Sodiumhydroxide (3.1 ml, 3.1 mM) was added, the mixture stirred at ambienttemperature for 30 minutes,. treated with acetic acid (0.3 ml), stirredfor a further 5 minutes, and then evaporated to dryness. The residue wastaken up in ethyl acetate (60 ml), washed with saturated aq. NaHCO₃ (60ml), brine, dried (MgSO₄) and evaporated, to give(2S,4S)-1-allyloxy-carbonyl-2-(3-allyloxycarbonyl-4-fluorophenylcarbamoyl)pyrrolidin-4-ylthiolas a gum (1.07 g, 89%). The crude material was used as such in the nextstage.

The thiol was condensed with carbapenem phosphate as in Example 1, andproduct purified by chromatography on silica, eluting with ethylacetate/dichloromethane (75:25) to give allyl (1R,5S,6S,8R,2S′,4S′)-2-(1-allyloxycarbonyl-2-(3-carboxy-4-fluorophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.24 (d, 3H); 1.35 (d, 3H); 2.40-2.80 (br, 2H); 3.24-3.28(m, 2H); 3.40-3.58 (br, 1H); 3.80 (dq, 1H); 3.99 (dd, 1H); 4.19-4.33 (m,2H); 4.53 (t, 1H); 4.59-4.77 (m, 4H); 4.77-4.88 (m, 2H); 5.17-5.50 (m,6H); 5.80-6.13 (m, 3H); 7.10 (dd, 1H); 7.82-7.95 (m, 1H); 8.00 (dd, 1H);8.70-9.20 (br s, 1H).

Ms (+ve FAB): 658 (MH)⁺

EXAMPLE 7

(1R,5S,6S,8R,2′S,4′S)-2-(2-(5-Carboxy-2-fluorophenylcarbamoyl)-pyrrolidin-4-yl-thio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid was prepared from the appropriate protected carbapenem as inExample 6. Nmr (DMSO-d₆+acetic acid-d₄): δ 1.30 (d, 6H); 1.84 (m, 1H);2.80-2.93 (m, 2H); 3.36 (dd, 1H); 3.58 (m, 1H); 3.66 (dd, 1H); 3.81 (m,1H); 4.09( dq, 1H); 4.21 (m, 1H); 4.31 (dd, 1H); 7.56 (dd, 1H); 8.07 (m,1H); 8.90 (d, 1H).

Ms (+ve FAB): 494 (MH)⁺, 516 (M+Na)⁺

The starting materials were prepared as follows:

Allyl 3-amino-4-fluorobenzoate

4-Fluoro-3-nitrobenzoic acid was allylated as in Example 6 to give allyl4-fluoro-3-nitro benzoate. Nmr (CDCl₃): δ 4.87 (dt, 2H); 5.48-5.32 (m,2H); 5.95-6.14 (m, 1H); 7.40 (dd, 1H); 8.30-8.38 (m, 1H); 8.76 (dd, 1H).

Ms (EI): 225 M⁺; (CI): 225 M⁺, 243 ((M+NH₄)⁺

The above was reduced essentially as in Example 2, except that methanolwas used as solvent to give allyl 3-amino-4-fluorobenzoate. Nmr (CDCl₃):δ 3.70 (br, 2H); 4.79 (dt, 2H); 5.25-5.44 (m, 2H); 5.96-6.09 (m, 1H);7.02 (dd, 1H); 7.41-7.54 (, 2H).

Ms (EI): 195 M⁺; (CI): 196 (MH)⁺

The above amine was condensed with the proline acid as in Example 1,purifying the product by chromatography on silica using ethylacetate/hexane (50:50) as eluent, to give(2S,4S)-1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-fluorophenylcarbaoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.33 (s, 3H); 2.62 (br, 2H); 3.40 (dd, 1H); 3.98-4.20 (m,2H); 4.60 (t, 1H); 4.67 (dt, 2H); 4.82 (dt, 2H); 5.2-5.5 (m, 4H).;5.8-6.15 (m, 2H); 7.16 (dd, 1H); 7.83 (ddd, 1H); 8.97 (dd, 1H); 9.27(br, 1H).

Ms (EI): 451 (MH)⁺; (CI): 451 (MH)⁺

The above thioacetate was deacetylated to thiol, condensed withcarbapenem phosphate as in Example 6, and the product purified bychromatography (eluting with ethyl acetate) to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-fluorophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (DMSO-d₆+acetic acid-d₄): δ 1.90 (m, part obscured, 1H); 2.78 (m,1H); 3.22 (dd, 1H); 3.26 (m, 1H); 3.52 (m, 1H); 3.83-4.03 (m, 2H); 4.09(dd, 1H); 4.22 (m, 1H); 4.53 (m, 1H); 4.53-4.65 (m, 4H); 4.80 (dt, 2H);5.05-5.43 (m, 6H); 5.73-5.98 (m, 2H); 5.98-6.09 (m, 1H); 7.38 (dd, 1H);7.78 (br s, 1H); 8.49 (m, 1H); 8.62 (d, 1H).

Ms (+ve FAB): 658 (MH)⁺, 680 (M+Na)⁺

EXAMPLE 8

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-2,4-difluorophenylcarbamoylpyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, was prepared from the appropriate protected carbapenem as inExample 7. Nmr: (DMSO-d₆+acetic acid-d₄): δ 1.29 (d, 6H); 1.94-1.99 (m,part obscured 1H); 2.81-3.07 (m, 1H); 3.05 (dd, 1H); 3.37 (dd, 1H); 3.56(m, 1H); 3.75 (dd, 1H); 3.92 (q, 1H); 4.10 (dq, 1H); 4.29-4.38 (m, 2H);7.22 (t, 1H); 8.07 (m, 1H).

Ms (+ve FAB): 512 (MH)⁺, 534 (M+Na)⁺

The starting materials were prepared as follows:

Allyl 3-amino-2,6-difluorobenzoate

2,6-Difluoro-3-nitrobenzoic acid was allylated as in Example 6 to giveallyl 2,6-difluoro-3-nitrobenzoate. Nmr (CDCl₃): 4.90 (dt, 2H);5.33-5.50 (m, 2H); 5.95-6.03 (m, 1H); 7.09-7.27 (ddd, 1H); 8.22-8.27(ddd, 1H).

Ms (EI): 244 (MH)⁺; (CI): 261 ((M+NH₄)⁺

The above ester was reduced as in Example 2, except that methanol wasused as the solvent, to give allyl 3-amino-2,6-difluorobenzoate. Nmr(CDCl₃): δ 4.86 (dt, 2H); 5.27-5.49 (m, 2H); 5.95-6.09 (m, 1H);6.77-6.86 (m, 2H).

MS (EI): 213 M⁺; (CI): 214 (MH)⁺

The above amine was condensed with the proline acid as in Example 1,purifying by chromatography and eluting with ethyl acetate/hexane(40:60), to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-2,4-difluorophenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.32 (s, 3H); 2.60 (br, 2H); 3.40 (dd, 1H); 3.95-4.19 (m,2H); 4.57 (t, 1H); 4.67 (dt, 2H); 4.86 (dt, 2H); 5.20-5.50 (m, 4H);5.81-6.13 (m, 2H); 6.95 (ddd, 1H); 8.4 (ddd, 1H); 9.2 (br, 1H).

MS (+ve FAB): 469 (MH)⁺

The above thioacetate was deacetylated to thiol, and condensed withcarbapenem phosphate as in Example 6. The product was purified bychromatography on silica, eluting with ethyl acetate, to give allyl(1R,5S,6S,8R,2S′,4S′)-2-(1-allyloxycarbonyl-2-(3-carboxy-2,4-difluorophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methyl-carbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.26 (d, 3H); 1.36 (d, 3H); 1.80 (d, 1H); 2.62 (br, 2H);3.19-3.36 (m, 2H); 3.44 (dd, 1H); 3.08 (q, 1H); 4.05 (dd, 1H); 4.19-4.33(m, 2H); 4.50-4.80 (m, 5H); 4.86 (dt, 2H); 5.17-5.50 (m, 6H); 5.82-6.10(m, 3H); 6.95 (dt, 1H); 8.38 (dt, 1H); 9.00 (br, 1H).

Ms (+ve FAB): 676 (MH)⁺, 698 (M+Na)⁺

EXAMPLE 9

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3,4-Dicarboxyphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, trisodium salt was prepared using the technique of Example 2except that a mixture of DMSO and THF was used. Nmr (DMSO-d₆+aceticacid-d₄): δ 1.18 (d, 6H); 1.84 (m, part obscured, 1H); 2.78 (m, 1H);3.02 (dd, 1H); 3.23 (dd, 1H); 3.40 (quintet, 1H); 3.58 (dd, 1H); 3.83(quintet, 1H); 4.00 (quintet, 1H); 4.20 (dd overlapping m, 2H); 7.92(dd, 1H); 8.17 (d, 1H); 8.33 (d, 1H).

Ms (+ve FAB): 542 (MH)⁺, (Na salt)⁺; 564 (Na₂ salt)⁺

The starting material was prepared as follows:

2-Carboxy-4-nitrobenzoic acid was allylated according to the procedureof Example 1 to give allyl 2-allyloxycarbonyl-4-nitrobenzoate. Nmr(CDCl₃): δ 4.85 (dt, 4H); 5.29-5.47 (m, 4H); 5.91-6.12 (m, 2H); 7.88 (d,1H); 8.38 (dd, 1H); 8.63 (d, 1H).

Reduction of the above nitro compound by the method of Example 2, andpurifying by medium pressure chromatography on silica using a gradientof dichloromethane/diethyl ether (100:0 to 90:10), gave allyl2-allyloxycarbonyl-4-amino-benzoate. Nmr (CDCl₃): δ 3.94 (br, 2H);4.71-4.80 (m, 4H); 5.22-5.42 (m, 4H); 5.88-6.10 (m, 2H); 6.69 (dd, 1H);6.75 (d, 1H); 7.74 (d, 1H).

The above amine was condensed with proline acid as Example 1, purifyingby medium pressure chromatography on silica using a gradient fromdichloromethane to 20% diethyl ether in dichloromethane, to give(2S,4S)-1-allyloxycarbonyl-2-(3,4-diallyloxycarbonylphenylcarbamoyl)-pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.32 (s, 3H); 2.59 (br, 2H); 3.37 (dd, 1H); 4.03(quintet, 1H); 4.12 (dd, 1H); 4.56 (t, 1H); 4.67 (d, 2H); 4.77 (t, 4H);5.25-5.42 (m, 6H); 5.84-6.11 (m, 3H); 7.79 (m, 3H); 9.52 (br, 1H).

The above thioacetate was deacetylated to thiol, which was condensedwithout further purification with carbapenem phosphate as Example 1,finally purifying by medium pressure chromatography on silica, usingdichloromethane/ethyl acetate 3:2 as eluant, to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3,4-diallyloxycarbonyl-phenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methyl-carbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.24 (d, 3H); 1.35 (d, 3H); 2.62 (br, 2H); 3.26 (ddoverlapping m, 2H); 3.47 (br, 1H); 3.81 (quintet, 1H); 3.97 (dd, 1H);4.19-4.29 (overlapping m, 2H); 4.53 (t, 1H); 4.62-4.82 (m, 8H);5.19-5.44 (m, 8H); 5.84-6.07 (m, 4H); 7.82 (s, -3H); 9.30 (br, 1H).

EXAMPLE 10

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-4-hydroxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid was prepared using the technique of Example 1, except that thecrude acid was of sufficient purity, and did not require chromatography.Nmr (DKSO-d₆+acetic acid-d₄): δ 1.17 (d, 6H); 1.95 (m, obscured, 1H);2.87 (m, obscured, 1H); 3.17 (dd, 1H); 3.25 (dd, 1H); 3.42 (dt, 1H);3.75 (dd, 1H); 3.99-4.05 (m, 2H); 4.22 (dd, 1H); 4.33 (t, 1H); 6.76 (d,1H); 7.56 (dd, 1H); 7.97 (d, 1H).

Ms (+ve FAB): 492 (MH)⁺, 514 (M+Na)⁺

The starting materials were prepared as follows:

2-Hydroxy-5-nitrobenzoic acid was allylated essentially as in Example 1,except that the final extraction solvent was diethyl ether, to giveallyl 2-allyloxy-5-nitrobenzoate. Nmr (CDCl₃): δ 4.82 (m, 4H); 5.26-5.55(m, 4H); 5.97-6.13 (m, 2H); 7.49 (d, 1H); 8.41 (dd, 1H); 8.52 (d, 1H).

Reduction of the above nitro compound by the method of Example 2, exceptthat the solvent was methanol, and NaHCO₃ solution was used to basify,gave allyl 2-allyloxy-5-aminobenzoate. Nmr (CDCl₃): δ 3.23 (br, 2H);4.53 (dt, 2H); 4.79 (d, 2H); 5.21-5.49 (m, 4H); 5.93-6.14 (m, 2H); 6.80(m, 2H); 7.16 (d, 1H).

The above amine was condensed with proline acid as Example 1, purifyingby chromatography using a gradient of dichloromethane/diethyl ether(100:0 to 85:15) to give(2S,4S)-1-allyloxycarbonyl-2-(4-allyloxy-3-allyloxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.33 (s, 3H); 2.56 (br, 2H); 3.39 (dd, 1H); 4.01(quintet, 1H); 4.13 (dd, 1H); 4.52 (t, 1H); 4.60 (dt, 2H) 4.66 (m, 2H);4.81 (dt, 2H); 5.23-5.51 (m, 6H); 5.85-6.13 (m, 3H); 6.91 (d, 1H); 7.76(dd, 1H); 7.81 (d, 1H); 8.97 (br, 1H).

The above thioacetate was deacetylated to thiol, and condensed withcarbapenem phosphate as Example 1, purifying by chromatography usinggradient elution from dichloromethane to ethyl acetate to give allyl(1R,5S, 6S,8R, 2′S,4′S)-2-(1-allyloxycarbonyl-2-(4-allyloxy-3-allyloxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDC13): δ 1.24 (d, 3H); 1.36 (d, 3H); 2.57 (br, 2H); 3.25, 3.28 (ddoverlapping quintet, 2H); 3.47 (br, 1H); 3.78 (quintet, 1H); 4.01 (dd,1H); 4.18-4.27 (dd overlapping m, 2H); 4.51 (t, 1H); 4.58-4.79 (m, 6H);4.79 (dt, 2H); 5.19-5.51 (m, 8H); 5.83-6.12 (m, 4H); 6.93 (d, 1H); 7.79(dd, 1H); 7.85 (d, 1H); 8.88 (br, 1H).

Ms (+ve FAB): 696 (MH)⁺, 718 (M+Na)⁺

EXAMPLE 11

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3,5-Dicarboxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid was prepared using the technique of Example 1, except that thecrude acid was of sufficient purity, and did not need chromatography.Nmr (DMSO-d₆+acetic acid-d₄): δ 1.15 (d, 6H); 1.77 (m, part obscured,1H); 2.69 (m, part obscured, 1H); 2.85 (m, part obscured, 1H); 3.19 (dd,1H); 3.33-3.51 (m, 2H); 3.71 (quintet, 1H); 3.94 (quintet, 1H); 4.03 (t,1H); 4.15 (dd, 1H); 8.18 (t, 1H); 8.45 (d, 2H).

Ms (+ve FAB): 520 (MH)⁺, 542 (M+Na)⁺

The starting materials were prepared as follows:

3-Carboxy-5-nitrobenzoic acid was allylated essentially as in Example 1to give allyl 3-allyloxycarbonyl-5-nitrobenzoate. Nmr (CDCl₃): δ4.89-4.93 (m, 4H); 5.33-5.50 (m, 4H); 5.97-6.17 (m, 2H); 9.00 (t, 1H);9.04 (d, 2H).

Reduction of the above nitro compound by the method of Example 2 gaveallyl 3-allyloxycarbonyl-5-aminobenzoate. Nmr (CDCl₃): δ 3.91 (br, 2H);4.80-4.84 (m, 4H); 5.26-5.45 (m, 4H); 5.96-6.11 (m, 2H); 7.53 (d, 2H);8.09 (t, 1H).

The above amine was condensed with proline acid as Example 1, to give(2S,4S)-1-allyloxycarbonyl-2-(3,5-diallyloxycarbonylphenyl-carbamoyl)pyrrolidin-4-yl-thioacetate.Nmr (CDCl₃): δ 2.33 (s, 3H); 2.60 (br, 2H); 3.40 (dd, 1H); 4.03(quintet, 1H); 4.14 (dd, 1H); 4.58 (t, 1H); 4.65-4.70 (m, 2H); 4.83-4.87(m, 4H); 5.24-5.47 (m, 6H); 5.84-6.16 (m, 3H); 8.39 (d, 2H); 8.45 (t,1H); 9.36 (br, 1H).

The above thioacetate was deacetylated to thiol, and condensed withcarbapenem phosphate as Example 1, to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3,5-diallyloxy-carbonylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.23 (d, 3H); 1.35 (d, 3H); 2.63 (br, 2H); 3.25, 3.29 (ddoverlapping m, 2H); 3.49 (br, 1H); 3.84 (quintet, 1H); 3.98 (dd, 1H);4.19-4.30 (m, 2H); 4.55 (t, 1H); 4.63-4.78 (m, 4H); 4.84 (d, 4H);5.19-5.45 (m, 8H); 5.84-6.12 (m, 4H); 8.46 (s, 3H); 9.18 (br, 1H).

Ms (+ve FAB): 724 (MH)⁺, 746 (M+Na)⁺

EXAMPLE 12

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid.

To a solution of 4-nitrobenzyl(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-allyloxy-carbonylphenylcarbamoyl)-1-(4-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate(10 g, 12 mM) and Heldrum's acid (5.2 g, 36 mM) in THF (70 ml), under anatmosphere of argon and with the exclusion of light, was addedtetrakis(triphenylphosphine)palladium (1.4 g, 1.2 mM). The mixture wasstirred at ambient temperature for 30 minutes. The mixture was dilutedwith ethyl acetate (230 ml) and added to a solution of sodiumbicarbonate (1.5 g) in distilled water (200 ml). 10% Pd-charcoal (4 g)was added and the mixture hydrogenated in an atmosphere of hydrogen for3 hours. The catalyst was filtered, the filtrate extracted with ethylacetate (2×100 ml) and ether (2×100 ml), and the aqueous layerconcentrated under reduced pressure to about 250 ml. This solution wassplit into two and each sample purified by passage through a 1 literHP20SS column using water as eluent. The pure fractions were collectedand freeze-dried giving the title compound as a pale yellow solid (3.5g). Nmr (DMSO-d₆+acetic acid-d₄, positions sensitive to exact solventratio): δ 1.19 (d, 6H); 1.94 (dt, 1H); 2.97 (dt, 1H); 3.13 (dd, 1H);3.25 (dd, 1H); 3.42 (dt, 1H); 3.68 (dd, 1H); 3.94 (quintet, 1H); 4.02(quintet, 1H); 4.22 (dd, 1H); 4.32 (t, 1H); 7.46 (t, 1H); 7.73 (dt; 1H);7.88 (dm, 1H); 8.27 (t, 1H).

Ms (+ve FAB): 498 (Na salt), 520 (di-Na salt).

The starting material was prepared as follows;

3-Nitrobenzoic acid (50 g, 0.3 N) was allylated by a similar method tothat described in Example 1. Solid K₂CO₃ (82.7 g, 0.6 M) was added tothe acid in dry DMF (700 ml) with stirring. There was a slight exothermand the mixture became thick. Allyl bromide (38.8 ml, 0.45 M) was addedover 30 minutes and the mixture was left stirring overnight. Afterfiltration through diatomaceous earth, the solution was evaporated todryness under reduced pressure and the residue partitioned between etherand aqueous NaHCO₃. The ether layer was washed with dilute HCl, brineand water, dried and evaporated giving a yellow oil (62 g). Nmr(DMSO-d₆): δ 4.85-4.92 (m, 2H); 5.27-5.5 (m, 2H); 5.97-6.2 (m, 1H); 7.85(t, 1H); 8.37-8.42 (dt, 1H); 8.48-8.54 (dq, 1H); 8.64 (t, 1H).

Without further purification this oil was reduced to allyl3-aminobenzoate (53 g) using stannous chloride and the method describedin example 6. Nmr (CDCl₃): δ 3.6 (broad, 2H); 4.77-4.82 (dt, 2H);5.24-5.44 (m, 2H); 5.96-6.1 (m, 1H); 6.83-6.88 (m, 1H); 7.17-7.25 (m,1H); 7.35-7.47 (m, 2H).

The above allyl 3-aminobenzoate (26.6 g, 0.15 M) was condensed with4-acetylthio-1-(4-nitrobenzyloxycarbonyl)-2-carboxy-pyrrolidine (55.2 g,0.15 M) by suspension in toluene (750 ml) and addition of EEDQ (44.5 g,0.18 M). The mixture was stirred overnight, diluted with EtOAc (2 l) andwashed with dilute HCl, water and brine. The EtOAc phase was dried andevaporated, and the residue recrystallised from ethanol giving(2S,4S)-4-acetylthio-1-(4-nitrobenzyloxycarbonyl)-2-(3-allyloxycarbonylphenylcarbamoyl)-pyrrolidine(67.7 g). Nmr (DMSO-d₆): δ 1.93 (quintet, 1H); 2.9 (m, 1H); 3.35 (m,1H); 3.91-4.14 (m, 2H); 4.49 (quintet, 1H); 4.81 (dd, 2H); 5.22 (dd,2H); 5.2-5.44 (m, 2H); 5.95-6.10 (m, 1H); 7.47 (d, 2H); 7.66 (t, 2H);7.8-7.93 (m, 2H); 8.18-8.3 (m, 2H); 10.31 (s, 1H).

The above thioacetate (52.7 g, 0.1 M) was converted to the thiol bydissolving it in degassed allyl alcohol (1 l) and adding aqueous NaOH(2M, 50 ml) at 0° C. After 3 hours aqueous HCl (2 H, 52.5 ml) was added,the solvent evaporated and the residue partitioned between EtOAc andbrine. The EtOAc phase was dried over MgSO₄, filtered and evaporated.The thiol was used without further purification.

A solution of 4-nitrobenzyl(1R,5R,6S,8R)-6-(1-hydroxyethyl)-1-methyl-2-diphenylphosphoryloxycarbapenem-3-carboxylate(59.68 g, 0.1 M) in acetonitrile (500 ml) and methylene chloride (120ml) was cooled to −15° C. and ethyl diisopropylamine (52.5 ml) was addedslowly.(2S,4S)-1-(4-Nitrobenzyloxycarbonyl)-2-(3-allyloxycarbonylphenyl-carbamoyl)pyrrolidin-4-ylthiol(0.1 M), in acetonitrile (400 ml) was added under argon and the mixtureleft overnight. The solvent was evaporated and the residue subjected tochromatography on silica, eluting with methylene chloride, EtOAc andacetonitrile, giving 4-nitrobenzyl(1R,5R,6S,8R,2′S,4′S)-2-(1-(4-nitrobenzyloxycarbonyl)-2-(3-allyloxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxy-ethyl)-1-methylcarbapenem-3-carboxylateas a yellow solid (52.6 g). Nmr (DMSO-d₆+CD₃OD): δ 1.26 (d, 3H); 1.35(d, 3H); 2.2-2.4 (m, 1H); 2.7-2.95 (m, 1H); 3.28-3.40 (m, 2H); 3.54-3.63(m, 1H); 3.8 (t, 1H); 4.01-4.1 (q, 1H); 4.21-4.33 (m, 2H); 4.61 (dd,1H); 4.73 (d, 2H); 5.17-5.45 (m, 6M); 5.93-6.11 (m, 1H); 7.37-8.22(complex pattern of doublets and double doublets, 12H).

EXAMPLE 13

The deprotection and hydrogenation were carried out by a similar methodto that described in example 12, except that 4-nitrobenzyl(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-allyloxycarbonyl-5-carbamoylphenyl-carbamoyl)-1-(4-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate(0.44 g) was used. After the hydrogenation, the aqueous layer was wasfreeze-dried, without HP20SS chromatography, to give(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-carbamoylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, as a pale yellow solid (125 mg). Nmr (DMSO-d₆+acetic acid-d₄): δ1.9 (d, 6H); 1.98-2.06 (m, 1H); 2.85-2.98 (m, 1H); 3.16-3.21 (m, 1H);3.26 (dd, 1H); 3.43 (quintet, 1H); 3.74 (dd, 1H); 3.91-3.97 (m, 1H); 4.0(t, 1H); 4.23 (dd, 1H); 4.39 (t, 1H); 8.24 (t, 1M); 8.33 (t, 1H); 8.44(t, 1H).

The starting material was prepared as follows:

5-Nitroisophthalic acid (5 g) was converted to the mono allyl esterusing one equivalent of allyl bromide (2 ml) using a similar method tothat described in example 1. The required acid (2.7 g) was extractedfrom the organic phase with aqueous NaHCO₃. The organic layer containedthe di-allyl ester (2.7 g). The mono acid,3-allyloxycarbonyl-5-nitrobenzoic acid, was obtained as a white solid.Nmr (CDC13): 4.87 (d, 2H); 5.3-5.5 (q, 2H); 5.97-6.15 (m, 1H); 9.01 (t,3H).

Ms (CI): 252 (MH)⁺

DCCI (1.3 g) was added to a solution of the above acid (1.5 g) andN-hydroxysuccinimide (0.76 g) in methylene chloride (50 ml) and themixture stirred at ambient temperature for 2 hours. A white solid wasfiltered and the solution evaporated to dryness. The active ester waspurified on silica gel eluting with methylene chloride then it wasdissolved in methylene chloride and treated with ammonia gas at 5° C.The white solid which precipitated was3-allyloxycarbonyl-5-nitrobenzamide (1.1 g). Nmr (DMSO-d₆): δ 4.9 (dt,2H); 4.88-4.93 (m, 2H); 6.03-6.11 (m, 1H); 7.83 (broad s, 1H); 8.55(broad s, 1H); 8.75 (t, 1H); 8.84 (t, 1H); 8.94 (t, 1H).

Ms (CI): 268 ((M+NH₄)⁺

3-Allyloxycarbonyl-5-nitrobenzamide (1 g) was reduced with SnCl₂ by asimilar method to the reduction in example 12, giving3-allyloxycarbonyl-5-aminobenzamide (0.5 g). Nmr (DMSO-d₆): 4.78 (dt,2H); 5.2-5.8 (broad, 2H); 5.26-5.45 (m, 2H); 5.91-6.13 (m, 1H); 7.22(broad s, 1H); 7.27 (t, 1H); 7.33 (t, 1H); 7.59 (s, 1H).

Ms (CI): 221 (MH)⁺

(2S,4S)-4-Acetylthio-1-(4-nitrobenzyloxycarbonyl)-2-carboxy-pyrrolidine(0.75g) was converted to the acid chloride and reacted with3-allyloxycarbonyl-5-aminobenzamide (0.45 g) by a similar method to thatdescribed in example 12. The crude product was subjected tochromatography on silica gel, eluting with EtOAc and giving4-acetylthio-1-(4-nitrobenzyloxycarbonyl)-2-(3-allyloxycarbonyl-5-carbamoylphenylcarbamoyl)pyrrolidine (0.58 g).Nmr (DMSO-d₆): δ 1.92-2.06 (m, 1H); 2.3 (s, 3H); 2.79-2.83 (m, 1H); 3.38(dd, 1H); 3.97-4.12 (m, 2H); 4.49 (dd, 1H); 4.83 (dt, 2H); 5.19 (dd,2H); 5.25-5.43 (m, 2H); 5.97-6.11 (m, 1H); 7.31 (broad s, 2H); 7.54 (d,2H); 8.04 (d, 2H); 8.12 (t, 1H); 8.26 (t, 1H); 8.31 (t, 1H); 10.0 (s,1H).

The thiol was generated from the above thioacetate by the methoddescribed in example 12.

A solution of 4-nitrobenzyl(1R,5R,6S,8R)-6-(1-hydroxyethyl)-1-methyl-2-diphenylphosphoryloxycarbapenem-3-carboxylate(0.6 g) and(2S,4S)-1-(4-nitrobenzyloxycarbonyl)-2-(3-allyloxycarbonyl-5-aminocarbamoylphenylcarbamoyl)pyrrolidin-4-ylthiol(0.48 g) in acetonitrile (20 ml) was reacted by a similar method to thatas described in example 12. Purification was by flash chromatography,eluting with EtOAc then 5% HeOH/EtOAc, giving 4-nitrobenzyl(1R,5R,6S,8R,2′S,4′S)-2-(1-(4-nitrobenzyloxycarbonyl)-2-(3-allyloxy-carbonyl-5-carbamoylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxy-ethyl)-1-methylcarbapenem-3-carboxylateas a yellow solid (0.44 g). Nmr (DMSO-d₆): δ 1.19-1.22 (d, 6H); 2.0-2.11(m, 1H); 2.8-2.92 (m, 1H); 3.32 (dd, 1H); 3.42-3.62 (m, 2H); 3.92-4.2(m, 2H); 4.31 (broad d, 1H); 4.53 (q, 1H); 4.84 (d, 2H); 4.98-5.46 (m,6H); 5.98-6.14 (m, 1H); 7.42-8.52 (complex pattern of doublets anddouble doublets, 11H).

EXAMPLE 14

The deprotection and hydrogenation was carried out by a similar methodto that described in example 12, except that p-nitrobenzyl(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-allyloxycarbonyl-6-carbamoylphenylcarbamoyl)-1-(4-nitrobenzyloxycarbonyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylatewas used. After the hydrogenation, the aqueous layer was freeze-dried togive(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-6-carbamoylphenyl-carbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid. Nmr (DMSO-d₆+acetic acid-d₄): δ 1.3-1.38 (2×d, 6H); 2.0-2.11 (m,1H); 2.85-2.96 (m, 1H); 3.09 (dd, 1H); 3.37 (dd, 1H); 3.53 (quintet,1H); 3.79 (dd, 1H); 3.94 (t, 1H); 4.16 (t, 1H); 4.33 (dd, 1H); 4.43 (t,1H); 7.87 (dd, 1H); 7.97 (d, 1H); 9.12 (d, 1H).

Ms (+ve FAB): 541 (Na salt), 563 (di-Na salt).

The starting material was prepared as follows:

Nitroterephthalic acid (6.33 g) in methylene chloride (75 ml) and THF(15 ml) was converted to the mono acid chloride using oxalyl chloride(2.63 ml), DMF (2.55 ml) and N-methylmorpholine (7.95 ml) at −10° C.After 1 hour the solvents were removed and, without furtherpurification, the product was dissolved in allyl alcohol (20 ml) and THF(10 ml) and stirred overnight at ambient temperature. The solvents wereremoved and the residue partitioned between EtOAc and aqueous NaHCO₃.Acidification of the NaHCO₃ solution and extraction with EtOAc gave theproduct, 4-allyloxycarbonyl-2-nitrobenzoic acid (6.8 g). Nmr (CDCl₃): δ4.89 (d, 2H); 5.31-5.49 (m, 2H); 5.95-6.15 (m, 1H); 7.94 (d, 1H); 8.36(dd, 1H); 8.54 (d, 1H). Without purification, this acid was converted bythe method above to the acid chloride which was dissolved in THF (100ml) at 0° C. Ammonia gas was bubbled into the solution until thereaction was completed. The solution was partitioned between EtOAc andwater, and the product from the organic fraction purified on silica,eluting with methylene chloride followed by EtOAc. The product (4 g),containing a small impurity, was reduced with SnCl₂ by a similar methodto that in example 12 giving 4-allyloxycarbonyl-2-aminobenzamide. Nmr(DMSO-d₆): δ 4.79 (d, 2H); 5.22-5.46 (m, 2H); 5.91-6.13 (m, 1H); 7.02(dd. 1H); 7.37 (d, 1H); 7.61 (d, 1H).(2S,4S)-4-Acetylthio-1-(4-nitrobenzyloxycarbonyl)-2-carboxy-pyrrolidine(1.58 g) was converted to the acid chloride by suspension in methylenechloride (25 ml) and addition of oxalyl chloride (1.52 ml). A few dropsof DMF were added. After 2 hours the solvents were evaporated and theacid chloride, dissolved in methylene chloride (10 ml), was added underargon to a solution of 4-allyloxycarbonyl-2-amino-benzamide (0.52 g) inTHF (10 ml) and methylene chloride (5 ml) containing N-methylmorpholine(0.38 ml) at 0° C. The reaction was left overnight and partitionedbetween methylene chloride and dilute aqueous HCl. The methylenechloride fraction was washed with water, brine and dried. Purificationwas by silica chromatography, eluting with increasing concentrations ofEtOAc in methylene chloride, giving(2S,4S)-4-acetylthio-1-(4-nitrobenzyloxycarbonyl)-2-(3-allyloxy-carbonyl-6-carbamoylphenylcarbamoyl)pyrrolidine(0.84 g). Nmr (CDCl₃): δ 2.25 (quintet, 1H);.2.29 (s, 3H); 2.83 (m, 1H);3.53 (dd, 1H); 4.02 (quintet, 1H); 4.2 (dd, 1H); 4.49 (dd, 1H); 4.85 (d,2H); 5.23-5.46 (m, 2H); 5.97-6.13 (m, 1H); 7.31-7.5 (broad, 2H); 7.57(d, 1H); 7.75 (dd, 1H); 7.81-7.95 (broad, 2H); 9.27 (d, 1H).

The thiol was generated from the above thioacetate by a similar methodto that described in example 12.

A solution of 4-nitrobenzyl(1R,5R,6S,8R)-6-(1-hydroxyethyl)-1-methyl-2-diphenylphosphoryloxycarbapenem-3-carboxylateand(2S,4S)-1-(4-nitrobenzyloxycarbonyl)-2-(2-carbamoyl-5-allyloxycarbonylphenyl-carbamoyl)pyrrolidin-4-ylthiolin acetonitrile (20 ml) were reacted as described in example 12, giving4-nitrobenzyl(1R,5R,6S,8R,2′S,4′S)-2-(1-(4-nitrobenzyloxycarbonyl)-2-(3-allyloxy-3-carbonyl-6-carbamoylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylateas a yellow foam (1.24 g). Nmr 1.23-1.35 (m, 6H); 2.2-2.38 (m, 1H);2.77-2.93 (m, 1H); 3.27 (dd, 1H); 3.28-3.42 (m 1H); 3.65-3.92 (m, 2H);4.17-4.35 (m, 3H); 4.53 (t, 1H); 4.83 (d, 2H); 4.92-5.44 (m, 6H);5.93-6.11 (m, 1H); 6.22-6.58 (broad, 2H); 7.35-8.23 (complex pattern ofdoublets and double doublets, 10H); 9.20 (d, 1H).

EXAMPLE 15

The deprotection and purification were carried out by a similar methodto that described in example 2 (chromatography of the final product wason an HP20SS column) except that allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(2-dimethylaminocarbonyl-3-allyloxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxy-ethyl)-1-methylcarbapenem-3-carboxylatewas used. The appropriate aqueous fractions from the column werefreeze-dried to give(1R,5S,6S,8R,2′S,4′S)-2-(2-(2-dimethylaminocarbonyl-3-carboxyphenyl-carbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid. Nmr (DMSO-d₆+acetic acid-d₄, mixture of rotamers): δ 1.17 (d, 6H);1.75 (m, 2H); 2.65,2.72 (2×s, overlapping m, 4H); 3.00,3.01 (2×s,overlapping m, 4H); 3.17 (dd, 1H); 3.41 (quintet, 1H); 3.55 (m, 2H);3.96 (m, 2H); 4.15 (dd, 1H); 7.43 (t, 1H); 7.68 (m, 1H); 8.20-8.40 (2×d,1H).

The starting material was prepared as follows:

2-Dimethylaminocarbonyl-3-nitrobenzoic acid (1 g) was to the allyl esterby a similar method to that described in example 1 for the formation ofallyl 3-allyloxy-5-aminobenzoate giving allyl2-dimethylaminocarbonyl-3-nitrobenzoate (0.88 g). Nmr (DMSO-d₆): δ 2.45(s, 3H); 2.97 (s, 3H); 4.79 (dd, 2H); 5.28-5.47 (m, 2H); 5.9-6.1 (m,1H); 7.81 (t, 1H); 8.28-8.4 (dq, 2H).

Ms (CI): 279 (MH)⁺

The allyl ester (0.44 g) was reduced with SnCl₂ by a similar method tothe reduction described in example 1, giving allyl2-dimethylaminocarbonyl-3-aminobenzoate (0.41 g) as a clear red oil. RNr(CDCl₃): δ 2.24 (s, 3H); 3.06 (s, 3H); 3.64 (broad, 2H); 4.68 (dd, 2H);5.17-5.37 (m, 2H): 5.82-6.02 (m, 1H); 6.83 (dd, 1H); 7.09-6.85 (m, 1H);7.37 (dd, 1H).

Ms (CI): 249 (MH)⁺

Allyl 2-dimethylaminocarbonyl-3-aminobenzoate (0.39 g, 1.4 mM) wascondensed with(2S,4S)-4-acetylthio-1-alloxycarbonyl-2-carboxypyrrolidine (0.42 g, 1.54mM) using a similar method to the EEDQ method described in example 12,giving(2S,4S)-1-allyloxycarbonyl-2-(2-dimethylaminocarbonyl-3-allyloxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthioacetate (0.84 g). Nmr (CDCl₃, mixture of rotamers): δ 2.32 (s, 3H);2.38 (br m, part obscured, 1H); 2.38 (br m, part obscured, 1H);2.74,2.76 (2×s overlapping br m, 4H); 3.11,3.14 (2×s, 3H); 3.36-3.45 (m,1H); 3.98-4.14 (m, 2H); 4.50 (dd, 1H); 4.56-4.79 (m, 4H); 5.16-5.44 (m,4H); 5.93-6.06 (m, 2H); 7.45 (t, 1H); 7.81-7.86 (m, 1H); 8.33-8.41 (2×d,1H); 8.60 (br, 1H).

Ms (CI): 504 (MH)⁺

The thiol was generated from the above thioacetate by a similar methodto that described in example 12.

A solution of allyl(1R,5R,6S,8R)-6-(1-hydroxyethyl)-1-methyl-2-diphenylphosphoryloxycarbapenem-3-carboxylate(0.73 g, 1.47 mN) and(2S,4S)-1-allyloxycarbonyl)-2-(2-dimethylaminocarbonyl-3-allyloxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthiolin acetonitrile (12 ml) were reacted by a similar method to thatdescribed in the preparation of protected carbapenem′ step in example 1,giving allyl(1R,5R,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(2-dimethylaminocarbonyl-3-allyloxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate as a white solid (0.63 g). Nmr(CDCl₃, mixture of rotamers): δ 1.24 (d, 3H); 1.35 (2×d, 3H); 1.98 (br,1H); 2.34 (br, 1H); 2.73 (s, 3H); 3.11,3.13 (2×s, 3H); 3.22-3.46 (m,3H); 3.66-3.85 (m, 1H); 4.00-4.26 (m, 3H); 4.50 (t, 1H); 4.62-4.80 (m,6H); 5.08-5.46 (m, 6H); 5.83-6.06 (m, 3H); 7.46 (td, 1H); 7.86 (d, 1H);8.29 (m, 1H); 8.55 (br, 1H).

Ms (CI): 711 (MH)⁺

EXAMPLE 16

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-5-acetamidophenylcarbamoyl)-pyrrolidin-4-ylthio1-6- (1-hydroxyethyl) -1-methylcarbapenem-3-carboxylic acid, dipotassiumsalt.

To a solution of 4-allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-(4-nitrobenzyloxycarbonyl)-2-(3-carboxy-5-acetamidophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate(0.4 g, 0.53 mmol) in DMF (20 ml) were added Heldrum's acid (0.2 g, 1.39mmol) and tetrakis triphenylphosphine palladium (40 mg, 0.035 mmol). Thereaction mixture was stirred for one hour, at ambient temperature. Asolution of 1M potassium phosphate buffer (20 ml) and zinc powder (0.5g) were added to the solution and after one hour, at ambienttemperature, the reaction mixture was filtered over diatomaceous earthand the pH of the filtrate adjusted to 7.5 with solid potassiumcarbonate. The solution was filtered, concentrated under reducedpressure and the resulting residue purified by reverse phasechromatography (Nucleosil C18), with water as eluant, to give, afterfreeze drying, the title compound (78 mg, 28%). Nmr (DMSO-d₆+aceticacid-d₄): δ 1.15 (d, 3H); 1.17 (d, 3H); 1.74 (m, 1H); 2.06 (s, 3H); 2.66(m, 1H); 2.84 (m, 1H); 3.20 (dd, 1H); 3.40 (m, 2H); 3.70 (m, 1H); 3.97(m, 2H); 4.16 (dd, 1H); 7.92 (s, 1H); 7.94 (s, 1H); 8.19 (s, 1H).

The starting material was prepared as follows:

A solution of(2S,4S)-4-acetylthio-2-carboxy-1-(4-nitro-benzyloxycarbonyl)pyrrolidine(1 g, 1.8 mmol) and EEDQ (0.53 g, 3 mmol) in chloroform (70 ml) wasstirred at ambient temperature for 1 hour. 3-Acetamido-5-aminobenzoicacid (0.53 g, 2.7 mmol) and diisopropylethylamine (0.7 ml, 4 mmol) werethen added to the reaction mixture, which was stirred for 2 hours, atambient temperature. After evaporation of the solvent, the crudecompound was purified by chromatography on HP20SS using methanol/water(80:20) as the eluant. Partial evaporation of the solvents andlyophilisation gave(2S,4S)-1-(4-nitrobenzyloxycarbonyl)-2-(3-carboxy-5-acetamidophenyl-carbamoyl)pyrrolidin-4-ylthioacetate(1 g, 67%).

(2S,4S)-1-(4-Nitrobenzyloxycarbonyl)-2-(3-carboxy-5-acetamidophenylcarbamoyl)pyrrolidin-4-ylthioacetatewas dissolved in a mixture of methanol (60 ml) and water (20 ml) and thepH of the solution was adjusted to 11 with a 1H aqueous solution ofNaOH. After 30 minutes at ambient temperature, the reaction mixture wasneutralised with methanol, evaporated and purified by chromatography onHP20SS, using methanol/water (80:20) as the eluant. Evaporation andlyophilisation gave(2S,4S)-1-(4-nitrobenzyloxycarbonyl)-2-(3-carboxy-5-acetamidophenylcarbamoyl)pyrrolidin-4-ylthiol(0.68 g. 74%). Nmr (DMSOd₆+acetic acid-d₄): δ 2.05 (s, 3H); 2.05 (m,1H); 2.75 (m, 1H); 3.20-3.80 (m, 2H); 4.00 (m, 1H); 4.42 (m, 1H); 5.40(br s, 2H); 7.45-8.30 (m, 7H).

To a solution of 4-nitrobenzyl (1R,5S,6S,8R)-6-(1-hydroxy-ethyl)-1-methyl-2-diphenylphosphoryloxycarbapenem-3-carboxylate(0.6 g, 1.2 mol) in DMF (12 ml) were added sequentiallydiisopropylethylamine (0.6 ml, 3.4 mmol),(2S,4S)-1-(4-nitrobenzyloxycarbonyl)-2-(3-carboxy-5-acetamidophenylcarbamoyl)pyrrolidin-4-ylthiol(0.6 g, 1.2 mmol), tri-n-butylphosphine (0.6 ml, 2.4 mmol) and water(0.1 ml, 5.5 mmol). The reaction mixture was stirred overnight, at 4°C., evaporated to dryness and the residue was purified by chromatographyon HP20SS resin, using acetonitrile/water (40:60) as the eluant.Evaporation and lyophilisation gave allyl(1R,5R,6S,8R,2S′,4′S)-2-(1-(4-nitrobenzyl-oxycarbonyl)-2-(3-carboxy-5-acetamidophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate(0.8 g, 88%). Nmr (DMSO-d₆+acetic acid-d₄): δ 1.15 (d, 3H); 1.17 (d,3H), 2.05 (s, 3H); 2.17 (m, 1H); 2.81 (m, 1H); 3.26 (dd, 1H); 3.36 (td,1H); 4.56-4.74 (m, 2H); 5.02-5.73 (m, 4H); 5.91 (m, 1H); 7.47 (d, 1H);7.68 (d, 1H); 7.85-7.99 (m, 3H); 8.20-8.29 (m, 2H).

EXAMPLE 17

(1R,5S,6S,8R,2′S,4′S)-2-(2-(4-Acetamido-3-carboxyphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, dipotassium salt.

The title compound was prepared from 4-allyl (1R,5S,6S,8R,2′S,4′S)-2-(1-(4-nitrobenzyloxycarbonyl)-2-(4-acetamido-3-carboxy-phenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate(diisopropylethylamine salt) using a similar method to that of example16. Nmr (DMSO-d₆+acetic acid-d₄): δ 1.14 (d, 3H); 1.16 (d, 3H); 1.75 (m,1H); 2.07 (s, 3H); 2.73 (m, 1H); 2.96 (dd, 1H); 3.21 (dd, 1H); 3.39 (m,1H); 3.54 (dd, 1H); 3.78 (m, 1H); 3.96 (m, 1H); 4.08 (t, 1H); 4.18 (dd,1H); 7.69 (dd, 1H); 8.18 (d, 1H); 8.42 (d, 1H).

MS (+ve FAB): 571 (MH)⁺, (K salt)

The starting material was prepared as follows:

(2S,4S)-1-(4-Nitrobenzyloxycarbonyl)-2-(4-acetamido-3-carboxyphenylcarbamoyl)pyrrolidin-4-ylthioacetatewas prepared from 2-acetamido-5-aminobenzoic acid using a similar methodto that of example 16. Nmr (DMSO-d₆+acetic acid-d₄): δ 1.95 (m, 1H);2.12 (s, 3H); 2.34 (s, 3H); 2.77 (m, 1H); 3.32 (m, 1H); 3.93-4.50 (m,3H); 5.04-5.32 (m, 2H); 7.47-8.42 (m, 7H).(2S,4S)-1-(4-Nitrobenzyloxycarbonyl)-2-(4-acetamido-3-carboxyphenylcarbamoyl)pyrrolidin-4-ylthiolwas prepared from(2S,4S)-1-(4-nitrobenzyloxycarbonyl)-2-(4-acetamido-3-carboxyphenylcarbamoyl)pyrrolidin-4-ylthioacetateusing a similar method to that of example 16.

Allyl(1R,5R,6S,8R,2′S,4′S)-2-(1-(4-nitrobenzyloxycarbonyl)-2-(4-acetamido-3-carboxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate(diisopropylethyl-amine salt) was prepared from(2S,4S)-1-(4-nitrobenzyloxycarbonyl)-2-(4-acetamido-3-carboxyphenylcarbamoyl)pyrrolidin-4-ylthioland allyl(1R,5R,6S,8R)-6-(1-hydroxyethyl)-1-methyl-2-diphenylphosphoryloxy-carbapenem-3-carboxylateusing a similar method to that of example 16. Nmr (CDCl₃)δ: 1.14-1.80(m, 23H); 2.17 (s, 3H); 2.67 (m, 1H); 3.00-3.30 (m, 3H); 3.35-3.90 (m,4H); 3.90-4.40 (m, 3H); 4.40-4.75 (m, 3H); 5.00-5.75 (m, 4H); 5.70-6.10(m, 1H); 7.38-8.65 (m, 7H).

EXAMPLE 18

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-5-methylsulphonamidophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid dipotassium salt.

The title compound was prepared from allyl(2R,5S,6S,8R,2′S,4′S)-2-(1-(4-nitrobenzyloxycarbonyl)-2-(3-carboxy-5-methylsulphonamidophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxy-ethyl)-1-methylcarbapenem 3-carboxylate using a similar method to that ofexample 16. Nmr (DMSOd₆+acetic acid-d₄): δ 1.16 (d, 3H); 1.17 (d, 3H);1.74 (m, 1H); 2.64 (m, 1H); 2.81 (dd, 1H); 3.01 (s, 3H); 3.20 (m, 1H);3.40 (m, 2H); 3.68 (m, 1H); 3.96 (m, 2H); 4.17 (dd, 1H); 7.54 (s, 1H);7.82 (s, 3H); 8.00 (s, 1H).

MS (+ve FAB): 607 (M+H)⁺ for monopotassium salt; 645 (M+H)⁺ fordipotassium salt.

The starting material was prepared as follows:

(2S,4S)-1-(4-(Nitrobenzyloxycarbonyl)-2-(3-carboxy-5-methyl-simlphonamidophenylcarbamoyl)pyrrolidin-4-ylthioaetate as prepared from3-amino-5-methylsulphonamidobenzoic acid using a similar method to thatof example 16. Nmr (DMSO-d₆+acetic acid-d₄): δ 2.00 (m, 1H); 2.30 (s,3H); 2.84 (m, 1H); 2.95 (s, 3H); 3.20-3.51 (m, 1H); 3.83-4.20 (m, 2H);4.30-4.58 (m, 1H); 35.20 (m, 2H); 7.48-8.22 ( d, 7H).

(2S,4S)-1- (4-Nitrobenzyloxycarbonyl) -2-(3-carboxy-5-methyl-sulphonamidophenylcarbamoyl) pyrrolidin-4-ylthiolwas prepared from (2S,4S) -1-(4-nitrobenzyloxycarbonyl)-2-(3-carboxy-5-methylsulphonamido-phenylcarbamoyl)pyrrolidin-4-ylthioacetateusing a similar method to that of example 16. Nmr (DMSO-d₆+aceticacid-d₄): δ 2.10 (m, 1H); 2.78 (m, 1H); 2.99 (s, 3H); 3.43 (m, 1H); 3.68(m, 1H); 4.05 (m, 1H); 4.42 (m, 1H); 5.13-5.32 (m, 3H); 7.50-8.82 (m,1H).

Allyl(1R,5R,6S,8R,2′S,4′S)-2-(1-(4-nitrobenzyloxycarbonyl)-2-(3-carboxy-5-methylsulphonamidophenylcarbanoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenes-3-carboxylatewas prepared from(2S,4S)-1-(4-nitrobenzyloxycarbonyl)-2-(3-carboxy-5-methylsulphonamido-phenylcarbamoyl)pyrrolidin-4-ylthioand allyl(1R,5R,6S,8R)-6-(1-hydroxethyl)-1-methyl-2-diphenylphosphoryloxycarbapenem-3-carboxylateusing a similar method to that of example 16. Nmr (DMSO-d₆+aceticacid-d₄): δ 1.20 (d, 3H); 1.22 (d, 3H); 1.96 (m, 1H); 2.83 (m, 1H); 3.00(s, 3H); 2.26-3.57 (m, 3H); 3.87-4.28 (m, 4H); 4.42-4.76 (m, 3H);5.08-5.42 (m, 4H); 5.92 (m, 1H); 7.22-8.22 (m, 7H).

EXAMPLE 19

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-5-sulphophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt.

To a stirred solution of 4-nitrobenzyl(1R,5S,6S,8R,2′S,4′S)-2-(1-(4-nitrobenzyloxycarbonyl)-2-(3-carboxy-5-sulphophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate(0.5 g, 0.56 mmol) in DMF (5 ml) and water (5 ml) was added a solutionof 1M sodium phosphate buffer (5 ml) followed by zinc powder (1 g). Thereaction mixture was stirred for 1 hour and the pH of the solution wasadjusted to 8 by adding a saturated aqueous solution of sodium hydrogencarbonate. After filtration over diatomaceous earth, the filtrate wasconcentrated and purified by subjecting to preparative chromatography(Nucleosil C-18), using water as the eluant. Concentration andlyophilisation of the required fractions gave the title compound (44 mg,12%). Nmr (DMSO-d₆+acetic acid-d₄): δ 1.15 (d, 3H); 1.16 (d, 3H); 1.78(m, 1H); 2.73 (m, 1H); 2.92 (m, 1H); 3.21 (dd, 1H); 3.40 (m, 1H); 3.48(m, 1H); 3.75 (m, 1H); 3.97 (m, 1H); 4.03 (m, 1H); 4.18 (m, 1H); 7.93(s, 1H); 8.11 (s, 1H); 8.29 (s, 1H).

MS (−ve FAB): 576 (M−H)⁻ for monosodium salt; 598 (M−H)⁻ for disodiumsalt.

The starting material was prepared as follows:

(2S,4S)-1-(4-Nitrobenzyloxycarbonyl)-2-(3-carboxy-5-sulpho-phenylcarbaxoyl)pyrrolidin-4-ylthioacetatewas prepared from 3-amino-5-sulphobenzoic acid using a similar method tothat of example 16. Nmr (DMSO-d₆+acetic acid-d₄): δ 1.97 (m, 1H); 2.30(s, 3H); 2.80 (m, 1H); 3.37 (m, 1H); 3.86-4.15 (m, 2H); 4.46 (m, 1H);5.05-5.28 (m, 2H); 7.46-9.25 (m, 7H).

(2S,4S)-1-(4-Nitrobenzyloxycarbonyl)-2-(3-carboxy-5-sulphophenylcarbamoyl)pyrrolidin-4-ylthiolwas prepared from(2S,4S)-1-(4-nitrobenzyloxycarbonyl)-2-(3-carboxy-5-sulphophenylcarbamoyl)pyrrolidin-4-ylthioacetateusing a similar method to that of example 16. Nmr (DRSO-d₆+aceticacid-d₄): δ 1.25 (d, 3H); 1.27 (d, 3H); 2.07 (m, 1H); 2.70 (m, 1H); 3.4(m, 1H); 3.67 (m, 1H); 3.99 (m, 1H); 4.49 (m, 1H); 5.07-5.30 (m, 2H);7.47-8.40 (m, 7H).

4-Nitrobenzyl (1R,5S,6S,8R,2′S,4′S)-2-(1-(4-nitrobenzyloxycarbonyl)-2-(3-carboxy-5-sulphophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylatewas prepared from(2S,4S)-1-(4-nitrobenzyloxycarbonyl)-2-(3-carboxy-5-sulphophenyl-carbamoyl)pyrrolidin-4-ylthioland 4-nitrobenzyl(1R,5R,6S,8R)-6-(1-hydroxyethyl)-1-methyl-2-diphenylphosphoryloxycarbapenem-3-carboxylateusing a similar method to that of example 1. Nmr (DMSO-d₆+aceticacid-d₄): δ 1.20 (d, 3H); 1.21 (d, 3H); 1.96 (m, 1H): 2.77 (m, 1H);3.18-3.47 (m, 2H); 3.66-4.90 (m, 6H); 5.04-5.50 (m, 4H); 7.30-8.35 (m,11H).

EXAMPLE 20

(5R,6S,8R,2′S,4′S)-2-(2-(3-Carboxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)carbapenem-3-carboxylicacid, disodium salt.

To a stirred solution of allyl(5R,6S,8R,4′S)-2-(1-allyloxycarbonyl)-2-(3-allyloxycarbonylphenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)carbapenem-3-carboxylate (0.185 g, 0.296 mmol) in CH₂Cl₂ (4 ml), atambient temperature, were added sequentially N-methylaniline (0.190 g,1.776 mmol), water (4 ml), sodium bicarbonate (100 mg) andtetrakis(triphenylphosphine)palladium (34 mg, 0.029 mmol). After 10minutes the aqueous phase was separated and injected onto a C18preparative HLPC column giving the title compound (43 mg, 27%). Nmr(DMSO-d₆+acetic acid-d₄): 1.16 (d, 3H); 1.81 (m, 1H); 2.64 (m, 1H); 2.84(m, 1H); 3.26 (m, 3H); 3.4 (m, 1H); 3.67 (m, 1H); 3.94 (m, 2H); 4.12 (m,1H); 7.42 (t, 1H); 7.65 (d, 1H); 7.83 (d, 1H); 8.27 (s, 1H).

The starting allyl(5R,6S,8R,2′S,4′S)-2-(1-allyloxy-carbonyl-2-(3-allyloxycarbonylphenylcarbamoyl)pyrrolidin-4-yl-thio)-6-(1-hydroxyethyl)carbapenem-3-carboxylatewas prepared in 67% yield using a similar procedure to that described inexample 1, by reacting (2S,4S)1-allyloxycarbonyl-2-(3-allyloxycarbonylphenyl-carbamoyl)pyrrolidine-4-thiol,described in example 4, with allyl(5R,6S,8R)-6-(1-hydroxyethyl)-2-diphenylphosphoryloxycarbapenem-3-carboxylate(EP-A-126780 and EP-A-208889).

EXAMPLE 21

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-5-cyanophenylcarbamoyl-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt

To a solution of allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxy-carbonyl-2-(3-allyloxycarbonyl-5-cyanophenylcarbamoyl)pyrrolidin-4-yl-thio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate(358 mg, 0.54 mM) and 2,2-dimethyl-1,3-dioxane-4,6-dione (388 mg, 2.7mM) in a mixture of DMF (8 ml) and THF (4 ml), under an argonatmosphere, was added tetrakis(triphenylphosphine)palladium (62 mg,0.054 mM). The solution was stirred, under argon with protection fromlight, for 1.75 hours, and solvent removed by evaporation. The residuewas dissolved in a mixture of THF (6 ml) and DMF (2 ml), and a solutionof sodium 2-ethylhexanoate (295 mg, 1.77 mM) in THF (4 ml) was added,followed by diethyl ether (20 ml). The resultant precipitate wascentrifuged, and supernatant removed. The product was washed twice byresuspension in a mixture of THF (4 ml) and diethyl ether (10 ml), thendiethyl ether (20 ml) followed by centrifugation. Crude product wasdissolved in water (20 ml) and the pH adjusted to 7.4 with NAHCO₃. Afterfiltration, the solution was chromatographed on HP20SS resin, andfractions combined as appropriate to give the title product (206 mg,70%). Nmr (DMSO-d₆+acetic acid-d₄): δ 1.17 (d, 6H); 1.95 (m, partobscured, 1H); 2.83 (m, 1H); 3.09 (dd, 1H); 3.25 (dd, 1H); 3.41(quintet, 1H); 3.64 (dd, 1H); 3.88 (quintet, 1H); 4.02 (quintet, 1H);4.22 (dd, 1H); 4.31 (t, 1H); 8.00 (s, 1H); 8.28 (t, 1H); 8.46 (t, 1H).

Ms (+ve FAB): 523 (MH)⁺, (Na salt)

The starting materials were prepared as follows:

3-Cyano-5-nitrobenzoic acid

3-Amino-5-nitrobenzoic acid (3.64 g, 20 mM) was dissolved inconcentrated hydrochloric acid (20 ml), diluted with water (75 ml),cooled to 0°, and added over 30 minutes to a solution of NANO₂ (1.38g,20 mM) in water (10 ml). The pH was adjusted to 6.2 with saturatedNa₂CO₃ solution. A mixture of CuSO₄.5H₂O (10 g, 42 mM) in water (40 ml)and KCN (10 g, 154 mM) in water (20 ml) was heated to 65°, the solutionof diazonium salt added over 15 minutes, and the mixture refluxed for 40minutes. After cooling, and acidifying with 2M hydrochloric acid, theorganics were extracted into ethyl acetate (2×200 ml). The combinedextracts were washed with aqueous NaH₂PO₄, water, brine, and dried overNa₂SO₄. Evaporation gave 3-cyano-5-nitrobenzoic acid (3.6 g, 94%).

Nmr (DMSO-d₆): δ 8.69 (t, 1H); 8.80 (t, 1H); 8.97 (t, 1H).

Ms (−ve FAB): 191 (M−H)⁻

Ir (nujol): ν 2220 cm⁻¹

3-Cyano-5-nitrobenzoic acid was allylated essentially as in Example 1,except that the chromatographic purification used a mixture ofpetrol/ethyl acetate (5:1), to give allyl 3-cyano-5-nitrobenzoate. Nmr(DMSO-d₆): δ 4.91 (dt, 2H); 5.39-5.53 (m, 2H); 5.99-6.19 (m, 1H); 8.78(t, 1H); 8.81 (t, 1H); 9.04 (t, 1H).

Ms (+ve FAB): 202 M⁺; 232 ((M+NH₄)⁺; (both for amino compound by ammoniareduction)

Reduction of the above nitro compound by the method of Example 1 gaveallyl 3-amino-5-cyanobenzoate, mp 112-113°. Nmr (DMSO-d₆): δ 4.79 (dt,2H); 5.25-5.45 (m, 2H); 5.94-6.13 (m overlapping br, 1H); 7.10 (t, 1H);7.37 (t, 1H); 7.48 (t, 1H).

Ms (+ve FAB): 202 M⁺; 232 (M+NH₄)⁺

The above amine was condensed with proline acid as Example 1, purifyingby chromatography using a gradient of dichloromethane/ethyl acetate(19:1 to 9:1) to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxy-carbonyl-5-cyanophenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.33 (s, 3H); 2.59 (br, 2H); 3.38 (dd, 1H); 3.97-4.17 (m,2H); 4.56 (t, 1H); 4.69 (d, 2H); 4.84 (d, 2H); 5.26-5.48 (m, 4H);5.85-6.14 (m, 2H); 8.03 (br s, 1H); 8.18 (t, 1H); 8.29 (br s, 1H); 9.69(br, 1H).

Ms (+ve FAB): 458 (MH)⁺; 480 (M+Na)⁺

The above thioacetate was deacetylated as Example 1, to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-cyanophenylcarbamoyl)pyrrolidin-4-ylthiol.Nmr (CDCl₃): δ 1.90 (d, 1H); 2.52 (br, 1H); 2.65 (br, 1H); 3.34-3.52 (m,2H); 4.07 (dd, 1H); 4.54 (t, 1H); 4.69 (d, 2H); 4.84 (d, 2H); 5.27-5.47(m, 4H); 5.87-6.11 (m, 2H); 8.01 (s, 1H); 8.21 (t, 1H); 8.28 (s, 1H);9.56 (br, 1H).

The above thiol was condensed with carbapenem phosphate as Example 1,purifying by chromatography using a gradient of dichloromethane/ethylacetate (3:2 to 2:3) to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-cyanophenylcarbamoyl)-pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-aethylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.27 (d, 3H); 1.35 (d, 3H); 2.67 (v br, 2H); 3.21-3.33(overlapping m, 2H); 3.53 (br, 1H); 3.83 (quintet, 1H); 3.93 (dd, 1H);4.20-4.31 (overlapping m, 2H); 4.54 (t, 1H); 4.63-4.86 (m, 6H);5.21-5.47 (m, 6H); 5.82-6.11 (m overlapping br, 3H); 8.05 (t, 1H); 8.33(br s, 1H); 8.37 (br s, 1H); 9.35 (br, 1H).

Ms (+ve FAB): 665 (MH)⁺; 687 (M+Na)⁺

EXAMPLE 22

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-5-methoxyphenyl-carbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 21. Nmr(DMSO-d₆+acetic acid-d₄): δ 1.17 (d, 3H); 1.18 (d, 3H); 1.83 (m, partobscured, 1H); 2.76 (quintet, 1H); 2.98 (dd, 1H); 3.22 (dd, 1H); 3.39(quintet, 1H); 3.56 (dd, 1H); 3.81 (s overlapping m, 4H); 4.00 (quintet,1H); 4.15,4.18 (t overlapping dd, 2H); 7.22 (t, 1H); 7.58 (t, 1H); 7.84(t, 1H).

Ms (−ve FAB): 505 (M−H)⁻, (Na salt)

The starting materials were prepared as follows:

3-Hydroxy-5-nitrobenzoic acid was methylated essentially as theallylation step of Example 1, except that the allyl bromide was replacedby dimethyl sulfate, and purification by chromatography was unnecessary,to give methyl 3-methoxy-5-nitrobenzoate. Nmr (CDCl₃): δ 3.94 (s, 3H);3.97 (s, 3H); 7.87 (t, 1H); 7.90 (t, 1H); 8.44 (t, 1H).

The above ester (3.45 g, 16 mM) was dissolved in THF (100 ml), treatedwith 1M NaOH (25 ml), and stirred at ambient temperature for 5 hours.After removal of the solvent, the residue was treated with water (50ml), acidified with 2M sulfuric acid, and extracted with ethyl acetate(3×60 ml). The combined organic extracts were washed with aqueousNaH₂PO₄, brine, and dried over MgSO₄. Evaporation gave3-methoxy-5-nitrobenzoic acid, which was allylated essentially as inExample 1, except that the chromatographic purification used a mixtureof petrol/ethyl acetate (6:1), to give allyl 3-methoxy-5-nitrobenzoate.Nmr (CDCl₃): δ 3.95 (s, 3H); 4.87 (dt, 2H); 5.31-5.48 (m, 2H); 5.95-6.15(m, 1H); 7.89 (t, 1H); 7.92 (t, 1H); 8.46 (t, 1H).

Ms (CI): 237 M⁺; 255 ((M+NH₄)⁺

Reduction of the above nitro compound by the method of Example 1 gaveallyl 3-amino-5-methoxybenzoate. Nmr (DMSO-d₆): δ 3.72 (s, 3H); 4.50 (vbr, 2H); 4.75 (dt, 2H); 5.25-5.43 (m, 2H); 5.95-6.11 (m, 1H); 6.47 (t,1H); 6.75 (t, 1H); 6.93 (t, 1H).

Ms (CI): 208 (MH)⁺

The above amine was condensed with proline acid as Example 1, purifyingby chromatography using a gradient of petrol/ethyl acetate (5:2 to 2:1)to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-methoxyphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.32 (s, 3H); 2.59 (br, 2H); 3.38 (dd, 1H); 3.85 (s, 3H);4.02 (quintet, 1H); 4.15 (dd, 1H); 4.55 (t, 1H); 4.68 (d, 2H); 4.81 (d,2H); 5.22-5.46 (m, 4H); 5.83-6.13 (m, 2H); 7.35 (t, 1H); 7.58 (br s,1H); 7.64 (t, 1H); 9.12 (br, 1H).

Ms (+ve FAB): 463 (MH)⁺; 485 (M+Na)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradient ofdichloromethane/ethyl acetate (60:40 to 45:55) to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyloxy-carbonyl-5-methoxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxy-ethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.25 (d, 3H); 1.37 (d, 3H); 2.64 (v br, 2H); 3.21-3.33(overlapping m, 2H); 3.48 (br, 1H); 3.80 (quintet, 1H); 3.85 (s, 3H);4.01 (dd, 1H); 4.19-4.29 (overlapping m, 2H); 4.53 (t, 1H); 4.62-4.83(m, 6H); 5.20-5.45 (m, 6H); 5.84-6.11 (m overlapping br, 3H); 7.25 (t,1H); 7.63 (m, 2H); 8.90 (br, 1H).

Ms (+ve FAB): 670 (MH)⁺; 692 (M+Na)⁺

EXAMPLE 23

(1R,5S,6S,8R,2′S,4′S)-2-(2-(5-Carboxy-2-methanesulphinylphenylcarbamoyl)pyrrolidin-4-yl-thio)-6-(1-hydroxyethyl)-1-methyl-carbapenem-3-carboxylicacid, disodium salt was prepared as a mixture of diastereoisomers at thesulfoxide centre, using the technique of Example 21. Nmr (DMSO-d₆+aceticacid-d₄): δ 1.18 (d, 6H); 1.87 (m, part obscured, 1H); 2.64-2.91, 2.82,2.84 (m overlpping 2×s, 5H); 3.21 (dd, 1H); 3.40 (quintet, 1H); 3.52(dd, 1H); 3.71 (quintet, 1H); 4.01 (quintet, 1H); 4.01 (m, 1H), 4.18(dd, 1H); 7.83 (t, 1H); 7.94 (td, 1H); 8.45 (d, 1H).

Ms (+ve FAB): 560 (MH)⁺, (Na salt); 582 (MH)⁺, (Na₂ salt)

The starting materials were prepared as follows:4-Methanesulphinyl-3-nitrobenzoic acid 4-Methylthio-3-nitrobenzoic acid(4.36 g, 20 mM) was dissolved in acetic acid (200 ml), and treated atambient temperature with H₂O₂ (2.5 ml, 30%, 22 mN). After stirring atambient temperature for 4 days, excess peroxide was decomposed withsodium metabisulfite, and solvent evaporated. The residue was purifiedby chromatography on silica, eluting with methanol, to give4-methanesulphinyl-3-nitrobenzoic acid (4.1 g, 89%), mp 238-2390. Nmr(DMSO-d₆): δ 2.90 (s, 3H); 8.29 (d, 1H); 8.56 (dd, 1H); 8.68 (d, 1H).

Ms (−ve FAB): 229 (M−H)⁻

The above acid was allylated essentially as in Example 1, except thatthe chromatographic purification was unnecessary, to give allyl4-methanesulphinyl-3-nitrobenzoate, up 119-1210. Nmr (DMSO-d₆): δ 2.91(s, 3H); 4.91 (dt, 2H); 5.29-5.50 (m, 2H); 5.99-6.18 (m, 1H); 8.34 (d,1H); 8.61 (dd, 1H); 8.69 (d, 1H).

Reduction of the above nitro compound by the method of Example 1 gaveallyl 3-amino-4-methanesulphinylbenzoate. Nmr (CDCl3): δ 2.92 (s, 3H);4.82 (dt, 2H); 5.17 (br, 2H); 5.28-5.46 (m, 2H); 5.96-6.11 (m, 1H); 7.30(dd, 1H); 7.41 (m, 2H).

Ms (EI): 223 (M−O)⁺; 239 M⁺

The above amine was condensed with proline acid as Example 1, purifyingby chromatography using a gradient of hexane/ethyl acetate (3:2 to 1:1)to give(2S,4S)-1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-methanesulphinylphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (DMSO-d₆ at 100°): δ 1.98 (m, 1H); 2.32 (s, 3H); 2.73 (s, 3H); 2.86(m, part obscured, 1H); 3.36 (m, 1H); 3.91-4.10 (overlapping m, 2H);4.48 (dd, 1H); 4.53 (m, 2H); 4.86 (d, 2H); 5.11-5.49 (m, 4H); 5.80-5.97(m, 1H); 6.01-6.15 (m, 1H); 8.09 (s, 2H); 8.35 (s, 1H).

Ms (+ve FAB): 495 (MH)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradientfrom ethyl acetate to ethyl acetate/isopropanol (98:2) to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-methanesulphinylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (DMSO-d₆, mixture of rotamers): δ 1.18 (d, 6H); 1.99 (br, 1H); 2.80(s overlapping m, 5H); 3.25 (solvent overlapping m, 2H); 3.54 (m, 1H);3.90-4.18 (m, 3H); 4.15 (dd, 1H); 4.55 (m, 4H); 4.85 (d, 2H); 5.06 (d,1H); 5.14-5.46 (m, 6M); 5.79-6.13 (m, 3H); 7.80-8.05(m, 2H); 8.12 (m,1H); 10.10 (m, 1H).

Ms (+ve FAB): 702 (KH)⁺

EXAMPLE 24

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-5-methanesulphonylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 21. Nmr(DMSO-d₆+acetic acid-d₄): δ 1.16 (d, 3H); 1.18 (d, 3H); 1.75 (quintet,1H); 2.63 (m, 1H); 2.79 (dd, 1H); 3.20 (s overlapping m, 4H); 3.38 (m,2H); 3.64 (quintet, 1H); 3.95 (m, 2H); 4.14 (dd, 1H); 8.09 (t, 1H); 8.44(m, 2H).

Ms (−ve FAB): 574 (M−H)⁻, (Na salt)

The starting materials were prepared as follows:

3-Methylthio-5-nitrobenzoic acid 3-Amino-5-nitrobenzoic acid (1.82 g, 10mM) was dissolved in concentrated sulphuric acid (1.9 ml), diluted withwater (10 ml), and cooled to 5°. A solution of NaNO₂ (0.7 g, 10 mM) inwater (3 ml) was added and the mixture stirred for 30 minutes. The coldsolution of diazonium salt was added to a slurry of thiomethylcopper(I)at 3°, and the mixture stirred for 45 minutes. Organics were extractedinto ethyl acetate (5×60 ml), and the combined organic layers washedwith aqueous NaH₂PO₄, water, brine, and dried over Na₂SO₄. Evaporationgave 3-methylthio-5-nitrobenzoic acid (1.77 g, 83%). Nmr (DMSO-d₆): δ2.63 (s, 3H); 8.10 (t, 1H); 8.21 (t, 1H); 8.32 (t, 1H); 13.68 (br, 1H).

Ms (−ve FAB): 213 (M−H)⁻

The above acid was allylated essentially as in Example 1, except thatthe chromatography eluant was a mixture of petrol/ethyl acetate (6:1),to give allyl 3-methylthio-5-nitrobenzoate. Nmr (CDCl₃): δ 2.60 (s, 3H);4.87 (dt, 2H); 5.32-5.49 (m, 2H); 5.93-6.07 (m, 1H); 8.20 (m, 2H); 8.57(t, 1H).

Ms (EI): 253 M⁺

Allyl 3-methylsulphonyl-5-nitrobenzoate

Allyl 3-methylthio-5-nitrobenzoate (1.12 g, 4.4 mM) was dissolved inmethanol (30 ml), and cooled to 2°. A solution of “potassiumperoxymonopersulfate” (2KHSO₅.KHSO₄.K₂SO₄, 8.13 g, 13.2 mM) in water (25ml) was added slowly, and stirring continued for 4 hours. The mixturewas diluted with water (60 ml) and extracted with ethyl acetate (3×100ml). The combined organic extracts were washed with water, brine, anddried over Na₂SO₄. Crude product was purified by chromatography onsilica using a gradient of petrol/ethyl acetate (3:1 to 2:1) to giveallyl 3-methylsulphonyl-5-nitrobenzoate (0.74 g, 59%). Nmr (DMSO-d₆): δ3.44 (s, 3H); 4.94 (dt, 2H); 5.32-5.53 (m, 2H); 6.01-6.25 (m, 1H); 8.78(t, 1H); 8.89 (t, 1H); 8.91 (t, 1H).

Reduction of the above nitro compound by the method of Example 1 gaveallyl 3-amino-5-methanesulphonylbenzoate. Nmr (DMSO-d₆): δ 3.15 (s, 3H);4.80 (dt, 2H); 5.26-5.47 (m, 2H); 5.95-6.15 (m overlapping br, 3H); 7.29(t, 1H); 7.47 (t, 1H); 7.51 (t, 1H).

The above amine was condensed with proline acid as Example 1, purifyingby chromatography using a gradient of hexane/ethyl acetate (2:1 to 1:1)to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-methanesulphonylphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.34 (s, 3H); 2.48 (m, 1H); 2.62 (m, 1H); 3.17 (s, 3H);3.41 (dd, 1H); 4.03 (quintet, 1H); 4.15 (dd, 1H); 4.58 (dd, 1H); 4.71(d, 2H); 4.84 (dt, 2H); 5.27-5.47 (m, 4H); 5.88-6.14 (m, 2H); 8.23 (brs, 2H); 8.37 (t, 1H); 9.69 (br, 1H).

Ms (+ve FAB): 511 (MH)⁺; 533 (M+Na)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradient ofdichloromethane/ethyl acetate (55:45 to 20:80) to give allyl(1R,5S,6S,8R,2′S,⁴′S)-2-(1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-methanesulphonylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.24 (d, 3H); 1.36 (d, 3H); 2.54 (br, 1H); 2.66 (br, 1H);3.12 (s, 3H); 3.19-3.22 (overlapping m, 2H); 3.54 (br, 1H); 3.87(quintet, 1H); 3.94 (dd, 1H); 4.25,4.29 (quintet overlapping dd, 2H);4.55 (t, 1H); 4.65-4.80 (m, 4H); 4.85 (d, 2H); 5.20-5.46 (m, 6H);5.86-6.12 (m overlapping br, 3H); 8.31 (br, 1H); 8.43 (br, 1H); 8.52(br, 1H); 9.40 (br, 1H).

Ms (+ve FAB): 718 (MH)⁺; 740 (M+Na)⁺

EXAMPLE 25

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-5-trifluoromethylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 2. Nmr(DMSO-d₆+acetic acid-d₄): δ 1.16 (d, 6H); 1.92 (m, part obscured, 1H);2.77 (m, part obscured, 1H); 3.00 (dd, 1H); 3.21 (dd, 1H); 3.37(quintet, 1H); 3.59 (quintet, 1H); 3.80 (quintet, 1H); 3.97 (quintet,1H); 4.20 (m, 2H); 7.91 (br s, 1H); 8.27 (br s, 1H); 8.44 (br s, 1H).

Ms (−ve FAB): 542 (M−H)⁻, (acid); 564 (M−H)⁻, (Na salt)

The starting materials were prepared as follows:

3-Nitro-5-trifluoromethylbenzoic acid was allylated essentially as inExample 1, except that the product was sufficiently pure for use withoutchromatography, to give allyl 3-nitro-5-trifluoromethylbenzoate. Nmr(DMSO-d₆): δ 4.91 (dt, 2H); 5.3 -5.51 (m, 2H); 5.99-6.20 (m, 1H); 8.58(br s, 1H); 8.77 (br s, 1H); 8.84 (t, 1H).

Ms (CI): 275 M⁺; 293 ((M+NH₄)⁺

Reduction of the above nitro compound by the method of Example 2 gaveallyl 3-amino-5-trifluoromethylbenzoate, sufficiently pure for usewithout chromatography. Nmr (DMSO-d₆): 4.78 (dt, 2H); 5.24-5.43 (m, 2H);5.93-6.13 (m, 1H); 7.08 (t, 1H); 7.27 (br s, 1H); 7.44 (t, 1H).

Ms (CI): 245 M⁺; 263 ((M+NH₄)⁺

The above amine was condensed with proline acid as Example 1, purifyingby chromatography using a gradient from dichloromethane todichloromethane/diethyl ether (9:1) to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-trifluoromethylphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.33 (s, 3H); 2.59 (m, 2H); 3.39 (dd, 1H); 4.04 (quintet,1H); 4.14 (dd, 1H); 4.58 (t, 1H); 4.62 (dt, 2H); 4.85 (dt, 2H);5.23-5.48 (m, 4H); 5.84-6.15 (m, 2H); 8.03 (br s, 1H); 8.23 (br s, 2H);9.60 (br, 1H).

Ms (+ve FAB): 501 (MH)⁺; 523 (M+Na)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradientfrom dichloromethane to ethyl acetate/dichloromethane (9:1) to giveallyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-trifluoromethylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (DMSO-d₆+acetic acid-d₄): δ 1.21 (d, 6H); 2.05 (br, 1H); 2.85 (br,1H); 3.29 (dd, 1H); 3.44 (dd, 1H); 3.51 (quintet, 1H); 3.93 (br, 1H);4.05-4.18 (m, 2H); 4.27 (dd, 1H); 4.43-4.71 (overlapping m, 5H); 4.85(d, 2H); 5.16-5.46 (m, 6H); 5.70-6.16 (m, 3H); 7.94 (br s, 1H); 8.37 (brs, 1H); 8.53 (br s, 1H).

Ms (+ve FAB): 708 (MH)⁺; 730 (M+Na)⁺

EXAMPLE 26

(1R,5S,6S,8R,2′S,4′S)-2-(2-(5-Carboxy-2-methoxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 1. Nmr(DMSO-d₆+acetic acid-d₄): δ 1.20 (d, 6H); 1.93 (m, part obscured, 1H);2.86 (quintet, 1H); 3.02 (dd, 1H); 3.26 (dd, 1H); 3.44 (quintet, 1H);3.68 (dd, 1H); 3.84 (quintet, 1H); 3.95 (s, 3H); 4.03 (quintet, 1H);4.22 (dd, 1H); 4.34 (t, 1H); 7.13 (d, 1H); 7.78 (dd, 1H); 8.25 (d, 1H).

Ms (+ve FAB): 528 (MH)⁺, (Na salt); 550 (MH)⁺, (Na₂ salt)

The starting materials were prepared as follows:

4-Methoxy-3-nitrobenzoic acid was allylated essentially as in Example 1,except that the product was sufficiently pure for use withoutchromatography, to give allyl 3-nitro-4-methoxybenzoate. Nmr (CDCl₃): δ4.03 (s, 3H); 4.83 (dt, 2H); 5.29-5.46 (m, 2H); 5.93-6.14 (m, 1H); 7.14(d, 1H); 8.24 (dd, 1H); 8.52 (d, 1H).

Ms (CI): 237 M⁺; 255 ((M+NH₄)

Reduction of the above nitro compound by the method of Example 2 gaveallyl 3-amino-4-methoxybenzoate sufficiently pure for use withoutchromatography. Nmr (CDCl₃): δ 3.72 (br, 2H); 3.90 (s, 3H); 4.77 (dt,2H); 5.24-5.43 (m, 2H); 5.95-6.10 (m, 1H); 6.79 (d, 1H); 7.41 (d, 1H);7.50 (dd, 1H).

Ms (CI): 208 (MH)⁺

The above amine was condensed with proline acid as Example 4, exceptthat the material was purified by chromatography on silica, using agradient from dichloromethane to dichloromethane/diethyl ether (4:1),giving(2S,4S)-1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-methoxyphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.33 (s, 3H); 2.51 (br, 1H); 2.66 (br, 1H); 3.41 (dd,1H); 3.93 (s, 3H); 4.01 (quintet, 1H); 4.17 (dd, 1H); 4.55 (t, 1H); 4.64(d, 2H); 4.80 (dt, 2H); 5.18-5.44 (m, 4H); 5.81-6.14 (m overlapping br,2H); 6.91 (d, 1H); 7.84 (dd, 1H); 8.90 (br, 1H); 9.01 (d, 1H).

Ms (+ve FAB): 463 (MH)⁺; 485 (M+Na)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradientfrom dichloromethane to ethyl acetate to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-methoxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.23 (d, 3H); 1.36 (d, 3H); 2.52 (br, 1H); 2.66 (br, 1H);3.22 (dd, 1H); 3.28 (quintet, 1H); 3.44 (dd, 1H); 3.83 (quintet, 1H);3.93 (s, 3H); 4.09 (m, 1H); 4.19-4.31 (overlapping m, 2H); 4.53 (t, 1H);4.65 (m, 4H); 4.81 (d, 2H); 5.19-5.45 (m, 6H); 5.83-6.11 (m, 3H); 6.91(d, 1H); 7.83 (dd, 1H); 8.79 (br, 1H); 9.04 (d, 1H).

Ms (+ve FAB): 670 (MH)⁺; 692 (M+Na)⁺

EXAMPLE 27

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-4-methoxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 2,except that the DMF was replaced by DMSO. Nmr (DMSO-d₆+acetic acid-d₄):δ 1.17 (d, 6H); 1.83 (m, part obscured, 1H); 2.75 (quintet, 1H); 2.97(dd, 1H); 3.23 (dd, 1H); 3.40 (quintet, 1H); 3.56 (dd, 1H); 3.82 (s+m,4H); 3.99 (quintet, 1H); 4.12 (t, 1H); 4.19 (dd, 1H); 7.09 (d, 1H); 7.75(dd, 1H); 7.95 (d, 1H).

Ms (+ve FAB): 528 (MH)⁺, (Na salt); 550 (MH)⁺, (Na₂ salt)

The starting materials were prepared as follows:

2-Hethoxy-5-nitrobenzoic acid was allylated essentially as in Example 1,except that the product was sufficiently pure for use withoutchromatography, to give allyl 2-methoxy-5-nitrobenzoate. Nmr (CDCl₃): δ4.03 (s, 3H); 4.85 (dt, 2H); 5.30-5.49 (m, 2H); 5.95-6.14 (m, 1H); 7.08(d, 1H); 8.48 (dd, 1H); 8.72 (d, 1H).

Ms (CI): 238 (MH)⁺; 255 ((M+NH₄)⁺

Reduction of the above nitro compound by the method of Example 2 gaveallyl 5-amino-2-methoxybenzoate sufficiently pure for use withoutchromatography. Nmr (CDCl₃): δ 3.39 (br, 2H); 3.83 (s, 3H); 4.80 (dt,2H); 5.23-5.47 (m, 2H); 5.94-6.13 (m, 1H); 6.83 (d, 2H); 7.18 (t, 1H).

Ms (CI): 208 (MH)⁺

The above amine was condensed with proline acid as Example 4, exceptthat the material was purified by chromatography on silica, using agradient from dichloromethane to dichloromethane/diethyl ether (4:1),giving(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-4-methoxyphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.32 (s, 3H); 2.58 (br, 2H); 3.39 (dd, 1H); 3.89 (s, 3H);4.02 (quintet, 1H); 4.13 (dd, 1H); 4.53 (t, 1H); 4.66 (dt, 2H); 4.80(dt, 2H); 5.23-5.48 (m, 4H); 5.84-6.13 (m, 2H); 6.94 (d, 1H); 7.80 (m,2H); 8.94 (br, 1H).

Ms (CI): 463 (MH)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradientfrom dichloromethane to ethyl acetate to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyloxycarbonyl-4-methoxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.25 (d, 3H); 1.36 (d, 3H); 2.65 (br, 2H); 3.25 (dd, 1H);3.28 (quintet, 1H); 3.47 (br, 1H); 3.79 (quintet, 1H); 3.89 (s, 3H);4.01 (dd, 1H); 4.18-4.29 (overlapping m, 2H); 4.51 (t, 1H); 4.66 (m,4H); 4.79 (dt, 2H); 5.19-5.46 (m, 6H); 5.84-6.11 (m, 3H); 6.95 (d, 1H);7.79-7.87 (m, 2H); 8.70 (br, 1H).

Ms (+ve FAB): 670 (MH)⁺; 692 (M+Na)⁺

EXAMPLE 28

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-2-methoxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 2,except that the DMF was replaced by DMSO. Nmr (DMSO-d₆+acetic acid-d₄):δ 1.18 (d, 6H); 1.85 (m, part obscured, 1H); 2.66-2.86 (overlapping m,2H); 3.21 (dd, 1H); 3.41 (quintet, 1H); 3.52-3.72 (overlapping m, 2H);3.82 (s, 3H); 3.99 (quintet, 1H); 4.08 (dd, 1H); 4.17 (dd, 1H); 7.17 (t,1H); 7.45 (dd, 1H); 8.41 (dd, 1H).

Ms (+ve FAB): 528 (MH)⁺, (Na salt); 550 (MH)⁺, (Na₂ salt)

The starting materials were prepared as follows:

2-Hydroxy-3-nitrobenzoic acid was methylated essentially as theallylation step of Example 1, except that the allyl bromide was replacedby methyl iodide, and purification by chromatography was unnecessary, togive methyl 2-methoxy-3-nitrobenzoate. Nmr (DMSO-d₆): δ 3.88 (s, 3H);3.90 (s, 3H); 7.44 (t, 1H); 8.04 (dd, 1H); 8.12 (dd, 1H).

Ms (CI): 212 (MH)⁺; 229 (M+NH₄)⁺

The above ester (3.45 g, 16 mM) was hydrolysed by essentially the methodof Example 22, except that the solvent was DMSO in place of THF, to give2-methoxy-3-nitrobenzoic acid. Nmr (DMSO-d₆): δ 3.89 (s, 3H); 7.40 (t,1H); 8.01 (dd, 1H); 8.06 (dd, 1H).

Ms (CI): 215 ((M+NH₄)⁺

The above nitro acid was allylated essentially as in Example 1, exceptthat the product was sufficiently pure for use without chromatography,to give allyl 2-methoxy-3-nitrobenzoate. Nmr (CDCl₃): δ 4.00 (s, 3H);4.86 (dt, 2H); 5.31-5.50 (m, 2H); 5.96-6.16 (m, 1H); 7.27 (d, 1H); 7.92(dd, 1H); 8.06 (dd, 1H).

Ms (CI): 238 (MH)⁺; 255 ((M+NH₄)⁺

Reduction of the above nitro compound by the method of Example 1 gaveallyl 3-amino-2-methoxybenzoate sufficiently pure for use withoutchromatography. Nmr (CDCl₃): δ 3.86 (s, 3H); 3.92 (br, 2H); 4.82 (dt,2H); 5.26-5.49 (m, 2H); 5.96-6.16 (m, 1H); 6.91 (dd, 1H); 7.00 (t, 1H);7.23 (dd, 1H).

Ms (CI): 208 (MH)⁺; 225 ((M+NH₄)⁺

The above amine was condensed with proline acid as Example 4, to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-2-methoxyphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.30 (s, 3H); 2.53 (br, 1H); 2.65 (br, 1H); 3.41 (dd,1H); 3.86 (s, 3H); 4.04 (quintet, 1H); 4.16 (dd, 1H); 4.58 (t, 1H); 4.66(d, 2H); 4.83 (dt, 2H); 5.20-5.47 (m, 4H); 5.83-6.13 (m overlapping br,2H); 7.16 (t, 1H); 7.60 (dd, 1H); 8.57 (dd, 1H); 9.15 (br, 1H).

Ms (+ve FAB): 463 (MH)⁺; 485 (M+Na)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradientfrom dichloromethane to ethyl acetate to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyloxycarbonyl-2-methoxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.24 (d, 3H); 1.35 (d, 3H); 2.53 (br, 1H); 2.68 (br, 1H);3.24 (dd, 1H); 3.-28 (quintet, 1H); 3.43 (br, 1H); 3.80 (quintet,1H);.3.83 (s, 3H); 4.12 (m, 1H); 4.19-4.29 (overlapping m, 2H); 4.57 (t,1H); 4.64 (m, 4H); 4.83 (d, 2H); 5.18-5.48 (m, 6M); 5.81-6.14 (m, 3H);7.17 (t, 1H); 7.61 (dd, 1H); 8.56 (dd, 1H); 9.02 (br, 1H).

Ms (+ve FAB): 670 (MH)⁺; 692 (M+Na)⁺

EXAMPLE 29

(1R,5S,6S,8R,2′S,4′S)-2-(2-(5-Carboxy-2-methylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 2,except that the DMF was replaced by DMSO. Nmr (DMSO-d₆+acetic acid-d₄):δ 1.19 (d, 6H); 1.88 (m, part obscured, 1H); 2.31 (s, 3H); 2.77 (dt,1H); 2.93 (dd, 1H).; 3.22 (dd, 1H); 3.42 (quintet, 1H); 3.57 (dd, 1H);3.77 (quintet, 1H); 4.01 (quintet, 1H); 4.16 (t, 1H); 4.19 (dd, 1H);7.35 (d, 1H); 7.69 (dd, 1H); 8.39 (d, 1H).

Ms (+ve FAB): 512 (MH)⁺, (Na salt); 534 (MH)⁺, (Na₂ salt); 556 (M+Na)⁺,(Na₂ salt);

The starting materials were prepared as follows:

4-Methyl-3-nitrobenzoic acid was allylated as in Example 1, except thatpurification by chromatography was unnecessary, to give allyl4-methyl-3-nitrobenzoate. Nmr (DMSO-d₆): δ 2.59 (s, 3H); 4.84 (dt, 2H);5.27-5.47 (m, 2H); 5.96-6.16 (m, 1H); 7.67 (d, 1H); 8.16 (dd, 1H); 8.44(d, 1H).

Ms (EI): 222 (MH)⁺

Reduction of the above nitro compound by the method of Example 2, exceptthat the solvent was methanol, gave allyl 3-amino-4-methylbenzoatesufficiently pure for use without chromatography. Nmr (DMSO-d₆): δ 2.10(s, 3H); 4.74 (dt, 2H); 5.15 (br, 2H); 5.22-5.43 (m, 2H); 5.93-6.12 (m,1H); 7.04 (d, 1H); 7.11 (dd, 1H); 7.28 (d, 1H).

Ms (CI): 192 (MH)⁺; 209 ((M+NH₄ )⁺

The above amine was condensed with proline acid as Example 4, purifyingby chromatography using a gradient from dichloromethane todichloromethane/diethyl ether (9:1), to give(2S,4S)-1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-methylphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (DMSO-d₆, mix of rotamers): δ 1.97 (quintet, 1H); 2.26 (s, 3H); 2.34(s, 3H); 2.80 (br, 1H); 3.30 (br, 1H); 3.93-4.08 (br m, 2H); 4.53 (br,3H); 4.80 (dt, 2H); 5.10-5.44 (m overlapping br, 4H); 5.78-6.13 (moverlapping br, 2H); 7.38 (d, 1H); 7.72 (dd, 1H); 7.94 (br, 0.5H); 8.01(br, 0.5H); 9.61 (br, 0.5H); 9.67 (br, 0.5H).

Ms (CI): 447 (MH)⁺; 464 ((M+NH₄)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradientfrom dichloromethane to dichloromethane/ethyl acetate (1:1), to giveallyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-methylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (DMSO-d₆+acetic acid-d₄, mix of rotamers): δ 1.16 (d, 3H); 1.18 (d,3H); 1.98 (quintet, 1H); 2.27 (s, 3H); 2.86 (br, 1H); 3.27 (dd, 1H);3.33 (t, 1H); 3.56 (quintet, 1H); 3.95 (quintet, 1H); 4.02 (dd, 1H);4.15 (quintet,, 1H); 4.27 (dd, 1H); 4.48-4.70 (overlapping m, 5H); 4.80(d, 2H); 5.10-5.45 (m overlapping br, 6H); 5.81-6.14 (m overlapping br,3H); 7.38 (d, 1H); 7.75 (dd, 1H); 8.01 (br, 0.5H); 8.07 (br, 0.5H); 9.60(br, 1H).

Ms (+ve FAB): 654 (MH)⁺; 676 (M+Na)⁺

EXAMPLE 30

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-4-methylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 2,except that the DMF was replaced by DMSO. Nmr (DMSO-d6+acetic acid-d₄):δ 1.15 (d, 3H); 1.16 (d, 3H); 1.78 (dt, 1H); 2.48 (s, 3H); 2.60 (dt,1H); 2.90 (dd, 1H); 3.21 (dd, 1H); 3.39 (quintet, 1H); 3.49 (dd, 1H);3.73 (quintet, 1H); 3.99 (quintet, 1H); 4.03 (t, 1H); 4.17 (dd, 1H);7.23 (d, 1H); 7.70 (dd, 1H); 8.12 (d, 1H).

Ms (+ve FAB): 512 (MH)⁺, (Na salt); 534 (MH)⁺, (Na₂ salt); 556 (M+Na)⁺,(Na₂ salt)

The starting materials were prepared as follows:

2-Hethyl-5-nitrobenzoic acid was allylated as in Example 1, except thatpurification by chromatography was unnecessary, to give allyl2-methyl-5-nitrobenzoate. Nmr (DMSO-d₆): δ 2.65 (s, 3H); 4.84 (dt, 2H);5.28-5.47 (m, 2H); 5.99-6.18 (m, 1H); 7.65 (d, 1H); 8.31 (dd, 1H); 8.57(d, 1H).

Ms (CI): 222 (MH)⁺; 099 ((M+NH)⁺

Reduction of the above nitro compound by the method of Example 2, exceptthat the solvent was methanol, gave allyl 5-amino-2-methylbenzoatesufficiently pure for use without chromatography. Nmr (DMSO-d₆): δ 2.33(s, 3H); 4.73 (dt, 2H); 5.18 (br, 2H); 5.23-5.44 (m, 2H); 5.93-6.12 (m,1H); 6.68 (dd, 1H); 6.95 (d, 1H); 7.12 (d, 1H).

Ms (CI): 192 (MH)⁺; 209 ((M+NH₄)⁺

The above amine was condensed with proline acid as Example 4, purifyingby chromatography using a gradient from dichloromethane todichloromethane/diethyl ether (9:1), to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-4-methylphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (DMSO-d₆, mix of rotamers): δ 1.91 (br m, 1H); 2.33 (s, 3H); 2.76(br m, 1H); 3.28 (s overlapping m, 4H); 4.00 (br m, 2H); 4.38 (t, 1H);4.51 (br, 2H); 4.78 (dt, 2H); 5.01-5.46 (m overlapping br, 4H);5.68-6.16 (m overlapping br, 2H); 7.27 (d, 1H); 7.72 (dd, 1H); 8.11 (br,0.5H); 8.05 (br, 0.5H); 10.17 (br, 1H).

Ms (+FAB): 447 (MH)⁺; 469 (M+Na)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradientfrom dichloromethane to dichloromethane/ethyl acetate (1:1), to giveallyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyl-oxycarbonyl-4-methylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (DMSO-d₆+acetic acid-d₄): δ 1.19 (d, 6H); 1.92 (br, part obscured,1H); 2.48 (s, 3H); 2.79 (br, 1H); 3.25 (dd, 1H); 3.32 (t, 1H); 3.54(quintet, 1H); 3.91 (br m, 1H); 4.01 (quintet, 1H); 4.12 (dd, 1H); 4.25(dd, 1H); 4.45 (m, 1H); 4.50-4.68 (m, 4H); 4.78 (dt, 2H); 5.18-5.45 (m,6H); 5.70-6.13 (m overlapping br, 3H); 7.25 (d, 1H); 7.76 (dd, 1H); 8.12(br m, 1H).

Ms (+ve FAB): 654 (MH)⁺; 676 (M+Na)⁺

EXAMPLE 31

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-5-methylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt

Allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-methylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate(3 g, 4.59 mm) and 2,2-dimethyl-1,3-dioxane-4,6-dione (3.97 g, 27.6 mM)were dissolved in a mixture of DMSO (15 ml) and THF (5 ml), under anargon atmosphere, and tetrakis(triphenylphosphine)palladium (531 mg,0.46 mN) was added. The solution was stirred, under argon withprotection from light, for 1 hour. A solution of sodium 2-ethylhexanoate(1.53 g, 9.22 mN) in THF (5 ml) was added, followed by THF (250 ml). Theresultant precipitate was filtered, under an argon blanket to excludemoisture, and washed successively with small portions of THF (twice),and diethyl ether. Crude product and NaHCO₃ (1.5 g) were dissolved inwater (100 ml), and the solution chromatographed on HP20SS resin using agradient elution from water to waterlacetonitrile (9:1). Appropriatefractions were combined and freeze-dried to give(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-methylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt. Nmr (DMSO-d₆+acetic acid-d₄): δ 1.13 (d, 3H); 1.15(d, 3H); 1.76 (dt, 1H); 2.32 (s, 3H); 2.68 (dt, 1H); 2.87 (dd, 1H); 3.18(dd, 1H); 3.36 (quintet, 1H); 3.45 (dd, 1H); 3.71 (quintet, 1H); 3.95(quintet, 1H); 4.02 (t, 1H); 4.14 (dd, 1H); 7.49 (s, 1H); 7.65 (s, 1H);8.05 (s, 1H).

Ms (+ve FAB): 512 (MH)⁺, (Na salt); 534 (MH)⁺, (Na₂ salt); 556 (M+Na)⁺,(Na₂ salt)

The starting materials were prepared as follows:

3-Methyl-5-nitrobenzoic acid

3,5-Dimethylnitrobenzene (30 g, 0.198 M) was heated with stirring to 80°in a mixture of pyridine′ (400 ml) and water (250 ml). KKnO₄ (62.7 g,0.396 M) was added in portions over 0.75 hours, and heating continued at85-90° for 1.75 hours. The hot solution was filtered through celite,washing with hot water (150 ml). The pink filtrates were decolourisedwith a few drops of sodium metabisulfite, and evaporated to dryness. Theresidue was dissolved in water (250 ml), and extracted with diethylether (2×90 ml). The aqueous layer was acidified (concentratedhydrochloric acid), and extracted with ethyl acetate (3×120 ml).Combined organic extracts were washed with NaH2PO4 solution, brine, anddried over MgSO₄. Crude product was eluted through a pad of silica,using a mixture of ethyl acetate/dichloromethane/acetic acid (25:25:1),to give 3-methyl-5-nitrobenzoic acid (14.5 g, 40%), mp 171-1720. Nmr(DMSO-d₆): δ 2.51 (s, 3H); 8.17 (s, 1H); 8.30 (t, 1H); 8.42 (t, 1H);13.58 (br, 1H).

Ms (CI): 181 (MH)⁺

3-Methyl-5-nitrobenzoic acid was allylated as in Example 1, except thatpurification by chromatography was unnecessary, to give allyl3-methyl-5-nitrobenzoate. Nmr (CDCl₃): δ 2.53 (s, 3H); 4.87 (dt, 2H);5.31-5.48 (m, 2H); 5.99-6.13 (m, 1H); 8.20 (s, 1H); 8.23 (s, 1H); 8.68(s, 1H).

Ms (CI): 222 (MH)⁺

Reduction of the above nitro compound by the method of Example 1, gaveallyl 5-amino-3-methylbenzoate sufficiently pure for use withoutchromatography. Nmr (CDCl₃): δ 2.30 (s, 3H); 3.46 (br, 2H); 4.78 (dt,2H); 5.23-5.45 (m, 2H); 5.93-6.12 (m, 1H); 6.68 (t, 1H); 7.17 (t, 1H);7.27 (t, 1H).

Ms (CI): 192 (MH)⁺; 220 (M+HC₂H₅)⁺

The above amine was condensed with proline acid as Example 4, purifyingby chromatography using hexane/ethyl acetate (3:1) as eluant, to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-methylphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.33 (s, 3H); 2.39 (s, 3H); 2.58 (br, 2H); 3.29 (dd, 1H);4.02 (quintet, 1H); 4.13 (dd, 1H); 4.56 (t, 1H); 4.68 (dm, 2H); 4.82(dt, 2H); 5.23-5.44 (m, 4H); 5.86-6.12 (m, 2H); 7.63 (s, 1H); 7.85 (s,1H); 7.85 (s, 1H); 9.09 (br, 1H).

Ms (+FAB): 447 (MH)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradient ofdichloromethane/ethyl acetate (3:2 to 2:3), to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-methylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.23 (d, 3H); 1.35 (d, 3H); 2.38 (s, 3H); 2.63 (br, 2H);3.23 (dd, 1H); 3.27 (quintet, 1H); 3.46 (br, 1H); 3.78 (quintet, 1H);4.00 (dd, 1H); 4.24 (overlapping m, 2H); 4.51 (t, 1H); 4.59-4.63 (i,4H); 4.79 (d, 2H); 5.17-5.42 (m, 6M); 5.82-6.09 (m, 3H); 7.61 (s, 1H);7.73 (s, 1H); 7.99 (s, 1H); 8.87 (br, 1H).

Ms (+ve FAB): 654 (MH)⁺

EXAMPLE 32

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-Carboxy-5-methoxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 2,except that the DMF was replaced by DMSO. Nmr (DMSO-d₆+acetic acid-d₄):δ 1.18 (d, 6H); 1.92 (br, part obscured, 1H); 2.81 (m, 1H); 3.03 (dd,1H); 3.23 (dd, 1H); 3.41 (quintet, 1H); 3.61 (m, 1H); 3.90 (soverlapping m, 4H); 4.00 (quintet, 1H); 4.21 (overlapping m, 2H); 8.25(t, 1H); 8.50 (m, 2H).

Ms (+ve FAB): 556 (MH)⁺, (Na salt); 578 (MH)⁺, (Na₂ salt)

The starting materials were prepared as follows:

3-Methoxycarbonyl-5-nitrobenzoic acid was allylated as in Example 1,except that purification by chromatography was unnecessary, to giveallyl 3-methoxycarbonyl-5-nitrobenzoate. Nmr (DMSO-d₆): δ 3.97 (s, 3H);4.91 (dt, 2H); 5.31-5.51 (m, 2H); 6.00-6.19 (m, 1H); 8.75 (t, 1H); 8.81(d, 2H).

Ms (EI): 265 M⁺

Reduction of the above nitro compound by the method of Example 2, exceptthat the solvent was methanol, gave allyl3-amino-5-methoxycarbonylbenzoate sufficiently pure for use withoutchromatography. Nmr (DMSO-d₆): δ 3.79 (br, 2H); 3.92 (s, 3H); 4.82 (dt,2H); 5.26-5.46 (m, 2H); 5.94-6.14 (m, 1H); 7.53 (m, 2H); 8.07 (t, 1H).

Ms (CI): 236 (MH)⁺; 253 ((M+NH⁴)⁺

The above amine was condensed with proline acid as Example 1, purifyingby chromatography using a gradient from dichloromethane todichloromethane/diethyl ether (9:1), to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-methoxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.33 (s, 3H); 2.60 (br, 2H); 3.40 (dd, 1H); 3.94 (s, 3H);4.04 (quintet, 1H); 4.14 (dd, 1H); 4.58 (t, 1H); 4.68 (dm, 2H); 4.85(dt, 2H); 5.25-5.47 (m, 4H); 5.85-6.16 (m, 2H); 8.36 (t, 1H); 8.43 (m,2H); 9.40 (br, 1H).

Ms (+FAB): 491 (MH)⁺; 513 (M+Na)⁺

(2S,4S)-1-Allyloxycarbonyl-2-(3-allyloxycarbonyl-5-methoxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthioacetate(1.2 g, 2.4 mM) was dissolved in THF (50 ml) under an argon atmosphere,and aqueous methylamine (33% w/v, 0.51 g, 5.4 mM) was added. Stirringwas continued for 3 hours, and solvent removed. The residue was treatedwith 2H hydrochloric acid and extracted with ethyl acetate. The organicsolution was washed with water, brine, aqueous NaHCO₃ and dried overMgSO₄. Removal of solvents gave(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-methoxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthiol(1.02 g, 93%). The thiol was condensed without further purification withcarbapenem phosphate as Example 1, purifying by chromatography using agradient from dichloromethane to dichloromethane/ethyl acetate (1:1), togive allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-methoxycarbonylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.24 (d, 3H); 1.37 (d, 3H); 2.64 (br, 2H); 3.26 (ddoverlapping quintet, 2H); 3.48 (br, 1H); 3.86 (quintet, 1H); 3.94 (soverlapping m, 4H); 4.25 (quintet, 1H); 4.29 (dd, 1H); 4.56 (t, 1H);4.69 (m, 4H); 4.86 (dm, 2H); 5.19-5.46 (m, 6H); 5.85-6.13 (m, 3H); 8.46(m, 3H); 9.18 (br, 1H).

Ms (+ve FAB): 698 (MH)⁺; 720 (M+Na)⁺

EXAMPLE 33

(1R,5S,6S,8R,2′S,4′S)-2-(2-(5-Carboxy-2,4-difluorophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 2. Nmr(DMSO-d₆+acetic acid-d₄): δ 1.20 (d, 6H); 1.96 (m, part obscured, 1H);2.81 (m, 1H); 3.14 (dd, 1H); 3.27 (dd, 1H); 3.43 (quintet, 1H); 3.73 (m,1H); 3.91 (quintet, 1H); 4.04 (quintet, 1H); 4.23 (dd, 1H); 4.43 (t,2H); 7.29 (t, 1H); 8.54 (t, 1H).

Ms (+ve FAB): 534 (MH)⁺, (Na salt); 556 (MH)⁺, (Na₂ salt)

The starting materials were prepared as follows:

2,4-Difluoro-5-nitrobenzoic acid

2,4-Difluorobenzoic acid (5 g, 0.031 M) was dissolved in concentratedsulfuric acid (30 ml), and cooled to 0°. The mixture was stirred, andfuming nitric acid (d 1.567 g/ml, 1.91 ml, 0.047 M) added dropwise,keeping the temperature below 5°. After stirring for 3 hours, the mixwas poured onto ice, and organics extracted into dichloromethane (2×75ml). The combined organic layers were washed with water, dried (MgSO₄),and evaporated to give 2,4-difluoro-5-nitrobenzoic acid (3.9 g, 61%).Nmr (DMSO-d₆): δ 7.18 (t, 1H); 8.88 (t, 1H); 9.93 (br, 1H).

Ms (−FAB): 202 (M−H)⁻

2,4-Difluoro-5-nitrobenzoic acid was allylated as in Example 1, exceptthat the reaction time was only 1.5 hours, the DMF was stirred overIR-120-H ion exchange resin before use, and purification bychromatography was unnecessary, to give allyl2,4-difluoro-5-nitrobenzoate. Nmr (CDCl₃): δ 4.88 (dt, 2H); 5.31-5.50(m, 2H); 5.93-6.13 (m, 1H); 7.13 (t, 1H); 8.80 (dd, 1H).

Ms (EI): 265 M⁺

Reduction of the above nitro compound by the method of Example 2, exceptthat the solvent was methanol, gave allyl 5-amino-2,4-difluorobenzoate,sufficiently pure for use without chromatography.

Nmr (CDCl₃): δ 3.61 (br, 2H); 4.81 (dt, 2H); 5.26-5.48 (m, 2H);5.92-6.12 (5, 1M); 6.83 (t, 1H); 7.38 (dd, 1H).

Ms (CI): 214 (MH)⁺; 231 ((M+NH⁴)⁺

The above amine was condensed with proline acid as Example 1, purifyingby chromatography using a gradient from dichloromethane todichloromethane/diethyl ether (9:1), to give(2S,4S)-1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2,4-difluorophenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.33 (s, 3H); 2.63 (br, 2H); 3.39 (dd, 1H); 4.04(quintet, 1H); 4.14 (dd, 1H); 4.59 (t, 1H); 4.66 (dt, 2H); 4.83 (dt,2H); 5.22-5.49 (m, 4H); 5.84-6.13 (m, 2H); 6.94 (t, 1H); 8.82 (t, 1H);9.22 (br, 1H).

Ms (+FAB): 469 (MH)⁺; 491 (M+Na)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradientfrom dichloromethane to dichloromethane/ethyl acetate (1:1), to giveallyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2,4-difluorophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.25 (d, 3H); 1.38 (d, 3H); 2.62 (br, 2H); 3.24 (ddoverlapping quintet, 2H); 3.45 (dd, 1H); 3.88 (quintet, 1H); 4.03 (dd,1H); 4.25 (quintet, 1H); 4.29 (dd, 1H); 4.57 (t, 1H); 4.68 (m, 4H); 4.82(dm, 2H); 5.21-5.48 (m, 6H); 5.85-6.10 (m, 3H); 6.94 (t, 1H); 8.85 (t,1H); 9.12 (br, 1H).

Ms (+ve FAB): 676 (MH)⁺; 698 (M+Na)⁺

EXAMPLE 34

(1R,5S,6S,8R,2′S,4′S)-2-(2-(5-Carboxy-2,4-dimethoxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 2. Nmr(DMSO-d₆+acetic acid-d₄): δ 1.20 (d, 6H); 1.95 (m, part obscured, 1H);2.87 (dt, 1H);.3.08 (dd, 1H); 3.26 (dd, 1H); 3.41 (quintet, 1H); 3.71(dd, 1H); 3.87 (quintet, part obscured, 1H); 3.91 (s, 3H); 3.96 (s, 3H);4.03 (quintet, 1H); 4.21 (dd, 1H); 4.39 (t, 2H); 6.76 (s, 1H); 8.44 (s,1H).

Ms (+ve FAB): 558 (MH)⁺, (Na salt); 580 (MH)⁺, (Na₂ salt)

The starting materials were prepared as follows:

2,4-Dimethoxy-5-nitrobenzoic acid

A solution of sodium methoxide was prepared by dissolving sodium metal(1.42 g, 0.062 M) in methanol (40 ml) with cooling. Allyl2,4-difluoro-5-nitrobenzoate (5 g, 0.021 M) was added and the mixturewas stirred for 2 hours. A solution of NaOH (2 g, 0.05 h) in water (10ml) was added, and the mixture stirred at ambient temperature for 16hours. Solvent was evaporated, the residue dissolved in water (50 ml),and extracted with diethyl ether (40 ml). The aqueous layer wasacidified with sulfuric acid, and the precipitate filtered and dried togive 2,4-dimethoxy-5-nitrobenzoic acid (4.23 g, 91%). Nmr (CDCl₃): δ4.07 (s, 3H); 4.16 (s, 3H); 6.62 (s, 1H); 8.81 (s, 1H).

Ms (CI): 228 (MH)⁺; 245 ((M+NH₄)⁺

2,4-Dimethoxy-5-nitrobenzoic acid was allylated as in Example 1, exceptthat purification by chromatography was unnecessary, to give allyl2,4-dimethoxy-5-nitrobenzoate. Nmr (CDCl₃): δ 4.02 (s, 3H); 4.04 (s,3H); 4.79 (dt, 2H); 5.26-5.46 (m, 2H); 5.93-6.13 (m, 1H); 6.54 (s, 1H);8.63 (s, 1H).

Ms (+FAB): 268 (MH)⁺; 290 (M+Na)⁺

Reduction of the above nitro compound by the method of Example 1, gaveallyl 5-amino-2,4-dimethoxybenzoate. Nmr (CDCl₃): δ 3.25 (br, 2H); 3.87(s, 3H); 3.90 (s, 3H); 4.76 (dt, 2H); 5.22-5.46 (m, 2H); 5.93-6.12 (m,1H); 6.47 (s, 1H); 7.29 (s, 1H).

Ms (CI): 238 (MH)⁺

The above amine was condensed with proline acid as Example 4, purifyingby chromatography using a gradient from dichloromethane todichloromethane/diethyl ether (4:1), to give(2S,4S)-1-allyloxycarbonyl-2-(5-1-allyloxycarbonyl-2,4-dimethoxyphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.32 (s, 3H); 2.49 (br, 1H); 2.64 (br, 1H); 3.40 (dd,1H); 3.91 (s, 3H); 3.93 (s, 3H); 4.00 (quintet, 1H); 4.17 (dd, 1H); 4.53(t, 1H); 4.64 (d, 2H); 4.77 (dt, 2H); 5.19-5.46 (m, 4H); 5.80-6.14 (moverlapping br, 2H); 6.49 (s, 1H); 8.69 (br, 1H); 8.81 (s, 1H).

Ms (+FAB): 493 (MH)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradientfrom dichloromethane to ethyl acetate, to give allyl (1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2,4-dimethoxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.22 (d, 3H); 1.36 (d, 3H); 2.48 (br, 1H); 2.65 (br, 1H);3.23 (dd, 1H); 3.28 (quintet, 1H); 3.43 (dd, 1H); 3.80 (quintet, 1H);3.91 (s, 3H); 3.92 (s, 3H); 4.09 (dd, 1H); 4.24 (quintet, 1H); 4.27 (dd,1H); 4.51 (t, 1H); 4.66 (m, 4H); 4.77 (dt, 2H); 5.20-5.45 (m, 6H);5.83-6.11 (m overlapping br, 3H); 6.49 (s, 1H); 8.45 (br, 1H); 8.82 (s,1H).

Ms (+ve FAB): 700 (MH)⁺

EXAMPLE 35

(1R,5S,6S,8R,2′S,4′S)-2-(2-(5-carboxy-2-cyanophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 2,except that the DMF was replaced by DMSO. Nmr (DMSO-d₆+acetic acid-d₄):δ 1.17 (d, 6H); 1.83 (m, part obscured, 1H); 2.62-2.79 (overlapping m,2H); 3.18 (dd, 1H); 3.41 (quintet, 1H); 3.66 (quintet, 1H); 3.98(quintet, 1H); 4.07 (dd, 1H); 4.17 (dd, 1H); 7.79 (m, 2H); 8.67 (s, 1H).

Ms (+ve FAB): 523 (MH)⁺, (Na salt); 545 (MH)⁺, (Na₂ salt)

The starting materials were prepared as follows:

4-Cyano-3-nitrobenzoic acid

4-Chloro-3-nitrobenzoic acid (5.84 g, 29 mM), cuprous cyanide (5.2 g, 58mM), cuprous chloride (0.96 g, 9.7 mM), and quinoline (6.9 ml, 58 mM)were heated under an argon atmosphere at 180′ for 3.5 hours. Aftercooling, the mixture was dissolved in concentrated hydrochloric acid (60ml), diluted with water (80 ml), and extracted with ethyl acetate (3×100ml). The combined organic layers were washed with aqueous NaH₂PO₄,brine, and dried over MgSO₄. Crude product was purified bychromatography on silica, eluting with a mixture ofdichloromethane/acetic acid (98:2), to give 4-cyano-3-nitrobenzoic acid(2.65 g, 48%). Nmr (DMSO-d₆): δ 8.31 (d, 1H); 8.41 (dd, 1H); 8.68 (d,1H).

Ms (EI): 192 M⁺

The above nitro acid was allylated essentially as in Example 1,purifying the crude product by chromatography on silica, using an eluantof hexane:ethyl acetate (6:1), to give allyl 4-cyano-3-nitrobenzoate.Nmr (CDCl₃): δ 4.93 (dt, 2H); 5.37-5.50 (m, 2H); 5.97-6.13 (m, 1H); 8.03(d, 1H); 8.46 (dd, 1H); 8.94 (d, 1H).

Ms (CI): 221 (MH)⁺; 250 ((M+NH₄)⁺

Reduction of the above nitro compound by the method of Example 1 gaveallyl 3-amino-4-carbamoylbenzoate, recrystallised from ethylacetate/petrol, mp 149-150°. Nmr (DMSO-d₆): δ 4.77 (dt, 2H); 5.25-5.43(m, 2H); 5.96-6.11 (m, 1H); 6.72 (br, 2H); 7.04 (dd, 1H); 7.25 (br, 1H);7.35 (d, 1H); 7.63 (d, 1H); 7.87 (br, 1H).

Ms (EI): 220 M⁺

The above amine was condensed with proline acid as Example 1, purifyingby chromatography using hexane/ethyl acetate (1:1) to give(2S,4S)-1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-carbamoylphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃, mix of rotamers): δ 2.25 (quintet, 1H); 2.28 (s, 3H); 2.82(br, 1H); 3.52 (dd, 1H); 4.13 (t, 1H); 4.20 (br m, 1H); 4.52 (dd, 1H);4.61 (br, 2H); 4.85 (d, 2H); 5.01-5.48 (m overlapping br, 4H); 5.66-6.12(m overlapping br, 2H); 6.55 (br, 1H); 6.89 (br, 1H); 7.64 (br m, 1H);7.79 (br m, 1H); 9.30 (br m, 1H); 11.68 (br, 0.5H); 12.06 (br, 0.5H).

Ms (+ve FAB): 476 (MH)⁺

(2S,4S)-1-Allyloxycarbonyl-2-(5-allyloxycarbonyl-2-cyanophenylcarbamoyl)pyrrolidin-4-ylthioacetate

DMF (0.2 ml, 2.5 mM) was dissolved in acetonitrile (10 ml), cooled to−5°, and treated with oxalyl chloride (0.2 ml, 2.3 mM). After stirringfor 30 minutes, a solution of(2S,4S)-1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-carbamoylphenylcarbamoyl)pyrrolidin-4-ylthioacetate(1 g, 2.1 mM) in acetonitrile (15 ml) was added, followed by pyridine(0.38 ml, 4.6 mM). After 15 minutes, the mixture was diluted with ethylacetate (200 ml), washed with hydrochloric acid (2H, 20 ml), water,aqueous NaHCO₃, and brine, and dried over Na₂SO₄. Crude product waspurified by chromatography using a gradient from dichloromethane toethyl acetate to give(2S,4S)-1-allyloxycarbonyl-2-(5-allyloxycarbonyl-2-cyanophenylcarbamoyl)pyrrolidin-4-ylthioacetate(0.9 g, 93%). Nmr (CDCl₃): δ 2.32 (s, 3H); 2.63 (br, 2H); 3.46 (dd, 1H);4.06 (quintet, 1H); 4.16 (dd, 1H); 4.64 (t, 1H); 4.71 (dt, 2H); 4.85(dt, 2H); 5.22-5.47 (m, 4H); 5.87-6.14 (m, 2H); 7.67 (d, 1H); 7.87 (dd,1H); 8.96 (d, 1H); 9.42 (br, 1H).

Ms (+ve FAB): 458 (MH)⁺; 480 (M+Na)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradientfrom dichloromethane to ethyl acetate to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyloxycarbonyl-2-methoxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.24 (d, 3H); 1.35 (d, 3H); 2.60 (br, 2H); 3.25 (ddoverlapping quintet, 2H); 3.52 (br, 1H); 3.88 (quintet, 1H); 4.02 (dd,1H); 4.25 (quintet, 1H); 4.28 (dd, 1H); 4.55-4.74 (m, 5H); 4.85 (dt,2H); 5.18-5.468 (m, 6H); 5.83-6.11 (m, 3H); 7.65 (dt, 1H); 7.87 (dd,1H); 9.00 (br s, 1H); 9.25 (br, 1H).

Ms (+ve FAB): 665 (MH)⁺; 687 (M+Na)⁺

EXAMPLE 36

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-5-fluorophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 2,except that the DMF was replaced by DMSO. Nmr (DMSO-d₆+acetic acid-d₄):δ 1.20 (d, 6H); 1.99 (dt, 1H); 2.91 (dt, 1H); 3.26 (dd overlapping m,3H); 3.42 (quintet, 1H); 3.74 (dd, 1H); 3.96 (quintet, 1H); 4.04(quintet, 1H); 4.22 (dd, 1H); 4.41 (t, 1H); 7.43 (dm, 1H); 7.81 (dd,1H); 8.00 (t, 1H).

Ms (+ve FAB): 516 (MH)⁺, (Na salt); 538 (MH)⁺, (Na₂ salt)

The starting materials were prepared as follows:

3-Fluoro-5-nitrobenzoic acid

A vigorously stirred slurry of nitrosonium tetrafluoroborate (3.53 g,30.2 mM) in acetonitrile (50 ml) under an atmosphere of argon was cooledin an ice bath, and 3-amino-5-nitrobenzoic acid (5.0 g, 27.5 mM) wasadded in three portions. The temperature was then allowed to rise toambient, and the mixture was stirred for 48 hours. 1,2-Dichlorobenzene(50 ml) was added, and acetonitrile distilled from the mixture atreduced pressure. The mixture was then heated to 170° for 30 minutes,when gas evolution had ended. After cooling, the mix was poured intodichloromethane (200 ml), and extracted with NaHCO₃ solution. After backwashing the aqueous phase with dichloromethane, it was acidified (2Mhydrochloric acid), and organics extracted with ethyl acetate (2×100ml). The combined organic layers were washed with brine, and dried overMgSO₄. Crude product was purified by chromatography on silica, elutingwith a gradient from dichloromethane/acetic acid (99:1) todichloromethane/isopropanol/acetic acid (80:20:1), to give3-fluoro-5-nitrobenzoic acid (3.26 g, 64%). Nmr (DMSO-d₆): δ 8.14 (dm,1H); 8.37 (dt, 1H); 8.46 (m, 1H).

Ms (EI): 185 M⁺

Allyl 3-fluoro-5-nitrobenzoate

3-Fluoro-5-nitrobenzoic acid (3 g, 16.2 mM), p-toluenesulfonic acid(1.54 g, 8.1 mM), and allyl alcohol (50 ml) were heated to reflux,passing the distillate through 3a molecular sieves, for 16 hours. Aftercooling, the mixture was neutralised with triethylamine, and solventremoved. The residue was dissolved in ethyl acetate, washed with 2Mhydrochloric acid, aqueous NaHCO₃, and brine, and dried over MgSO₄.Crude product was purified by chromatography on silica, using a gradientelution from dichloromethane to ethyl acetate/dichloromethane (3:1), togive allyl 3-fluoro-5-nitrobenzoate. Nmr (CDCl₃): δ 4.89 (dt, 2H);5.33-5.49 (m, 2H); 5.95-6.15 (m, 1H); 8.11 (m, 2H); 8.70 (t, 1H).

Ms (CI): 226 (MH)⁺; 253 ((M+C₂H₅)⁺

Reduction of the above nitro compound by the method of Example 1 gaveallyl 3-amino-5-fluorobenzoate. Nmr (CDCl₃): δ 3.89 (br, 2H); 4.79 (dt,2H); 5.25-5.45 (m, 2H); 5.92-6.12 (m, 1H); 6.54 (dt, 1H); 7.07-7.15 (m,2H).

Ms (CI): 196 (MH)⁺

The above amine was condensed with proline acid as Example 1, purifyingby chromatography using a gradient from dichloromethane todichloromethane/diethyl ether (85:13) to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-fluorophenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃): δ 2.33 (s, 3H); 2.59 (br, 2H); 3.48 (dd, 1H); 4.03(quintet, 1H); 4.13 (dd, 1H); 4.56 (t, 1H); 4.68 (dt, 2H); 4.82 (dt,2H); 5.25-5.46 (m, 4H); 5.86-6.11 (m, 2H); 6.47 (dm, 1H); 7.72 (t, 1H);7.87 (dt m, 1H); 9.38 (br, 1H).

Ms (+ve FAB): 451 (MH)⁺; 473 (M+Na)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradientfrom dichloromethane to ethyl acetate to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyloxycarbonyl-5-fluorophenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃): δ 1.24 (d, 3H); 1.36 (d, 3H); 2.63 (br, 2H); 3.26 (dd, 1H);3.29 (quintet, 1H); 3.48 (br, 1H); 3.81 (quintet, 1H); 3.97 (dd, 1H);4.27 (dd overlapping m, 2H); 4.54 (t, 1H); 4.62-4.76 (m, 4H); 4.81 (dt,2H); 5.20-5.46 (m, 6H); 5.85-6.10 (m overlapping br, 3H); 7.48 (dt, 1H);7.83 (br s, 1H); 7.88 (dt, 1H); 9.18 (br, 1H).

Ms (+ve FAB): 658 (MH)⁺; 680 (M+Na)⁺

EXAMPLE 37

(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxy-N′-methylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid, disodium salt was prepared using the technique of Example 2,except that the DMF was replaced by DMSO, and product was purified bychromatography on a CHP20P column, eluting with water. Nmr(DMSO-d₆+acetic acid-d₄, run at 50°): δ 1.12 (d, 3H); 1.18 (d, 3H); 1.67(br, 1H); 2.13 (br, 1H); 3.02.(dd, 1H); 3.17 (dd, 1H); 3.28 (soverlapping a, 5H); 3.65 (br, 1H); 3.98 (quintet overlapping m, 2H);4.12 (dd, 1H); 7.58 (d, 1H); 7.91 (d, 1H); 7.98 (m, 1H).

Ms (+ve FAB): 512 (MH)⁺, (Na salt); 534 (MH)⁺, (Na₂ salt)

The starting materials were prepared as follows:

Allyl 3-methylaminobenzoate

Allyl 3-aminobenzoate (5 g, 28.2 mN) was dissolved in triethylorthoformate (50 ml) and trifluoroacetic acid (5 drops) was added. Thesolution was stirred and refluxed up through 3a molecular sieves for 5hours. Solvent was removed, and the residue dissolved in ethanol (50ml), followed by the addition of acetic acid (8.08 ml) and sodiumcyanoborohydride (6.85 g, 0.108 M) in several portions. The mixture wasstirred at ambient temperature for 16 hours, and solvent removed. Theresidue was dissolved in diethyl ether, washed with water, brine, anddried over MgSO₄. Crude product was purified by chromatography onsilica, eluting with a gradient from dichloromethane todichloromethane/ethyl acetate (95:5), to give allyl3-methylaminobenzoate (0.93 g, 17%). Nmr (CDCl₃): δ 2.88 (s, 3H); 4.81(dt, 2H); 5.23-5.45 (m, 2H); 5.94-6.13 (m, 1H); 6.83 (dd, 1H); 7.25 (dd,1H); 7.33 (t, 1H); 7.43 (dm, 1H).

Ms (CI): 192 (MH)⁺

The above amine was condensed with proline acid as Example 1, purifyingby chromatography using a gradient from dichloromethane todichloromethane/diethyl ether (4:1) to give(2S,4S)-1-allyloxycarbonyl-2-(3-allyloxycarbonyl-N′-methylphenylcarbamoyl)pyrrolidin-4-ylthioacetate.Nmr (CDCl₃, mixture of rotamers): δ 1.93 (m, 1H); 2.32 (s, 3H); 2.48 (m,part obscured, 1H); 3.28, 3.31 (2×s, 3H); 3.40 (quintet, 1H); 3.76 (m,1H); 4.01 (m, 1H); 4.24 (m, 1H); 4.50-4.74 (m, 2H); 4.86 (d, 2H);5.18-5.48 (m, 4H); 5.84-6.13 (m, 2H); 7.38-7.68 (m, 2H); 7.90-8.11 (m,2H).

Ms (+ve FAB): 447 (MH)⁺; 469 (M+Na)⁺

The above thioacetate was deacetylated and condensed with carbapenemphosphate as Example 1, purifying by chromatography using a gradientfrom dichloromethane to ethyl acetate to give allyl(1R,5S,6S,8R,2′S,4′S)-2-(1-allyloxycarbonyl-2-(3-allyloxycarbonyl-N′-methylphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylate.Nmr (CDCl₃, mixture of rotamers): δ 1.20 (2×d, 3H); 1.34 (2×d, 3H); 1.87(br, 1H); 2.30 (br, 1H); 3.29, 3.31 (2×s, overlapping m, 6H); 4.05-4.30(m, 4H); 4.50-4.81 (m, 5H); 4.96 (d, 2H); 5.20-5.48 (m, 6H); 5.85-6.13(m, 3H); 7.39-7.68 (m, 2H); 7.91-8.11 (m, 2H).

Ms (+ve FAB): 654 (MH)⁺; 676 (M+Na)⁺

What is claimed is:
 1. A dry-blended pharmaceutical compositioncomprising an alkali metal carbonate or bicarbonate and a compound ofthe formula (I)

or a pharmaceutically acceptable salt or in vivo hydrolysable esterthereof wherein: R¹ is 1-hydroxyethyl, 1-fluoroethyl or hydroxymethyl;R²is hydrogen or C₁₋₄alkyl; R³ is hydrogen or C₁₋₄alkyl; R⁴ and R⁵ arethe same or different and are selected from hydrogen, halo, cyano,C₁₋₄alkyl, nitro, hydroxy, carboxy, C₁₋₄alkoxy, C₁₋₄alkoxycarbonyl,aminosulphonyl, C₁₋₄alkylaminosulphonyl, di-C₁₋₄-alkylaminosulphonyl,carbamoyl, C₁₋₄alkylcarbamoyl, di-C₁₋₄alkylcarbamoyl, trifluoromethyl,sulphonic acid, amino, C₁₋₄alkylamino, di-C₁₋₄alkylamino,C₁₋₄alkanoylamino, C₁₋₄alkanoyl(N—C₁₋₄alkyl)amino,C₁₋₄alkanesulphonamido and —S(O)_(n)C₁₋₄alkyl and wherein n is zero, oneor two; with the proviso that there is no hydroxy or carboxy substituentin a position ortho to the link to —NR³— and that overall the compoundof formula (I) is acidic.
 2. The dry-blended pharmaceutical compositionaccording to claim 1, wherein R¹ is 1-hydroxyethyl.
 3. The dry-blendedpharmaceutical composition according to claim 1 or claim 2, wherein R²is hydrogen or methyl.
 4. The dry-blended pharmaceutical compositionaccording to claim 1 or claim 2, wherein R² is methyl.
 5. Thedry-blended pharmaceutical composition according to claim 1 or claim 2,wherein R³ is hydrogen.
 6. The dry-blended pharmaceutical compositionaccording to claim 1 or claim 2, wherein the compound has the structureof formula (IV):

wherein R³, R⁴ and R⁵ are as hereinbefore defined.
 7. The dry-blendedpharmaceutical composition according to claim 6 wherein R⁴ and R⁵ arethe same or different and selected from hydrogen, fluoro, chloro,hydroxy, carboxy, cyano, nitro, methyl, ethyl, methoxy, ethoxy,methoxycarbonyl, carbamoyl, methylcarbamoyl, dimethylcarbamoyl,trifluoromethyl, sulphonic acid, methylsulphinyl, methylsulphonyl,methanesulphonamido or acetamido.
 8. The dry-blended pharmaceuticalcomposition according to claim 6 wherein at least one of R⁴ and R⁵ inthe compound of formula (IV) is hydrogen.
 9. The dry-blendedpharmaceutical composition according to claim 7 wherein at least one ofR⁴ and R⁵ in the compound of formula (IV) is hydrogen.
 10. Thedry-blended pharmaceutical composition according to claim 6 wherein R⁴is hydrogen, carboxy, fluoro, chloro, methyl, methoxy, cyano, sulphonicacid or methoxycarbonyl and R⁵ is hydrogen.
 11. The dry-blendedpharmaceutical composition according to claim 1, wherein the compound is(1R,5S,6S,8R,2′S,4′S)-2-(2-(3-carboxyphenylcarbamoyl)pyrrolidin-4-ylthio)-6-(1-hydroxyethyl)-1-methylcarbapenem-3-carboxylicacid or a pharmaceutically acceptable salt or in vivo hydrolysable esterthereof.